000062
10.3205/000062
urn:nbn:de:0183-0000623
Research Article
Detection of HER-2/neu, c-myc amplification and p53 inactivation by FISH in Egyptian patients with breast cancer
Nachweis von HER-2/neu, c-myc-Amplifikation und p53-Inaktivierung durch FISH bei ägyptischen Patienten mit Brustkrebs
Ismail
Ismail
Manal F.
MF
Department of Biochemistry, Faculty of Pharmacy, Cairo University, Cairo, Egypt
author
Aly
Aly
Magdy Sayed
MS
Dr.
Biological Science Department, Faculty of Science, King Khaled University, P.O. Box 9004 Abha Saudi Arabia, Tel.: +966507531152, Fax: +9662290165Department of Zoology, Faculty of Science, Beni Sweef University, Beni Sweef, Egypt
magdyaly@yahoo.com
author
Khaled
Khaled
Hussein M.
HM
Department of Clinical Oncology, National Cancer Institute, Cairo University, Cairo, Egypt
author
Mohamed
Mohamed
Hanaa M.
HM
Department of Zoology, Faculty of Science, Beni Sweef University, Beni Sweef, Egypt
author
German Medical Science GMS Publishing House
Düsseldorf
610
breast cancer
fluorescent in situ hybridization (FISH)
genetic alterations
HER-2/neu
p53
c-myc
Brustkrebs
Fluoreszenz-in-situ-Hybridisierung (FISH)
genetische Veränderungen
HER-2/neu
p53
c-myc
20090304
20090427
20090506
engl
1612-3174
7
GMS German Medical Science
GMS Ger Med Sci
03
Brustkrebs ist weltweit die häufigste Ursache für krebsbedingte Todesfälle bei Frauen. Der klinische Verlauf bei dieser Erkrankung ist sehr unterschiedlich, und Kliniker sind bemüht, prognostische Parameter zu finden, die für jeden Patienten zuverlässig den Verlauf voraussagen und eine geeignete adjuvante Therapie vorgeben können. Die Amplifikation der zwei Onkogene HER-2/neu und c-myc und die Inaktivierung des Tumorsuppressorgens p53 treten bei Brustkrebs häufig auf. Das Ziel dieser Studie war es, mit dem Verfahren der Fluoreszenz-in-situ-Hybridisierung (FISH) die Amplifikation der beiden Onkogene HER-2/neu und c-myc sowie die Inaktivierung des Tumorsuppressorgens p53 zu messen und diese molekularbiologischen Marker mit den gebräuchlichen klinischen und pathologischen Angaben zu korrelieren.
Die Studie wurde an 34 Gewebeproben, die von 33 Frauen und einem Mann mit Brustkrebs stammen, und 17 Proben, die von 16 Frauen und einem Mann mit benignen Brustgewebeveränderungen gewonnen wurden, durchgeführt. Die Ergebnisse zeigen, dass die Werte von HER-2/neu, c-myc und p53 in der Krebsgruppe signifikant verändert waren im Vergleich zu der Gruppe mit gutartigen Tumoren. Bei der Beziehung der Gehalte der molekularen Marker zu den klinisch-pathologischen Befunden waren die Befunde bei p53 signifikant mit dem Alter der Patientin korreliert. Die diagnostische Sensitivität der untersuchten Marker war signifikant korreliert mit der Größe der Tumoren. Im Hinblick auf das Tumorgrading zeigten HER-2/neu und p53 eine signifikante Zunahme bei low-grade-Tumoren, während c-myc eine signifikante Zunahme bei high-grade-Tumoren aufwies. Bezüglich des Disease-Staging waren HER-2/neu und c-myc die einzigen Tumormarker, die einen signifikanten Anstieg im letzten Stadium der Erkrankung aufwiesen. p53 und HER-2/neu waren signifikant assoziiert mit positivem Lymphknotenstatus. Eine signifikante Korrelation bestand bei den Werten der drei Biomarker untereinander.
Zusammenfassend kann die Bestimmung von HER-2/neu, c-myc und p53 in den Gewebeproben die Patienten in verschiedene Risikogruppen einteilen.
Breast cancer is a leading cause of cancer-related deaths in women worldwide. The clinical course of this disease is highly variable and clinicians continuously search for prognostic parameters that can accurately predict prognosis, and indicate a suitable adjuvant therapy for each patient. Amplification of the two oncogenes HER-2/neu and c-myc and inactivation of the tumor suppressor gene p53 are frequently encountered in breast carcinomas. The purpose of this study was to use the fluorescence in situ hybridization (FISH) for the assessment of HER-2/neu and c-myc amplification and p53 inactivation and to relate these molecular markers with the commonly used clinical and pathological factors. The study was conducted on 34 tissue samples obtained from 33 females and 1 male with breast carcinomas and 17 samples obtained from 16 females and 1 male with benign breast lesions. Results revealed that the level of HER-2/neu, c-myc and p53 in the malignant group was significantly increased as compared to the benign group. On relating the level of the molecular markers to clinicopathological factors, p53 was significantly associated with increased patient’s age. The sensitivity of the investigated markers significantly increased with larger tumor size. Concerning tumor grade, HER-2/neu and p53 showed a significant increase in low-grade tumors whereas c-myc showed a highly significant increase in high-grade tumors. With regard to disease staging, HER-2/neu and c-myc were the only markers that showed significant increase at late stages of disease. p53 and HER-2/neu were significantly associated with positive lymph nodal status. A significant correlation was obtained between the levels of the three biomarkers to each other. Conclusively, the combination of HER-2/neu, c-myc and p53 can stratify patients into different risk groups.
Introduction
Breast cancer ranks first among cancers affecting woman throughout the world and its marked impact is not res<![CDATA[tricte</PlainText></TextGroup>d to Western industrialized societies <TextLink reference="49"></TextLink>. Latest estimates suggest that more than 1,050,000 new breast cancer cases occur worldwide annually, with nearly 580,000 cases in developed countries and the remainder in developing countries <TextLink reference="17"></TextLink>. Breast cancer is the most common malignancy among Egyptian females accounting about 37.6% of all malignancies in women <TextLink reference="36"></TextLink>. Over the three years 2000–2002, 1831 breast cancer cases were registered; 1810 females and 21 males with an average of 603 cases of female breast cancer per year. </Pgraph>
<Pgraph>The progression of malignant tumors is a consequence of multiple alterations of the genome including either activation of oncogenes such as HER-2/neu and c-myc or inactivation of tumor suppressor genes such as p53 <TextLink reference="54"></TextLink>, <TextLink reference="48"></TextLink>, <TextLink reference="8"></TextLink>.</Pgraph>
<Pgraph>HER-2/neu oncogene is a member of the human epidermal growth receptor family <TextLink reference="12"></TextLink>. The gene product is a protein with extracellular and intracellular domain with tyrosine kinase activity involved in signal transduction of cell growth and development <TextLink reference="23"></TextLink>. Amplification of the HER-2/neu receptor tyrosine kinase has been implicated in the pathogenesis and aggressive behavior of approximately 25% of invasive human breast cancers. It has been suggested that aberrant HER-2 signaling contributes to tumor initiation and disease progression <TextLink reference="52"></TextLink>. Determination of HER-2/neu in serum of patients with metastatic breast cancer has proved clinical utility since it correlates with protein expression in primary tumors <TextLink reference="53"></TextLink>.</Pgraph>
<Pgraph>The c-myc oncogene has been shown to be amplified and/or overexpressed in many types of cancer. The frequencies of its amplification, mRNA and protein overexpression in breast cancer vary between 1%–94%, 22%–95%, and 50%–100%, respectively <TextLink reference="26"></TextLink>, <TextLink reference="6"></TextLink>. c-myc protein is a product of c-myc proto-oncogene and exerts diverse effects on cell behavior. c-myc, being a transcription factor, plays important role in cell growth and differentiation and was found to be associated with a more aggressive tumor and a poorer patient survival <TextLink reference="33"></TextLink>.</Pgraph>
<Pgraph>The human p53 gene encodes a 53 KD nuclear phosphoprotein consisting of 393 amino acids <TextLink reference="30"></TextLink>. Wild-type p53 protein blocks the entry of cells into S phases whereby allowing DNA repair to occur before replicating the genome. p53 can also induce apoptosis when DNA repair fails <TextLink reference="25"></TextLink>. However, not only mutant p53 cannot prevent the propagation of genetically damaged cells but also can inhibit apoptosis <TextLink reference="27"></TextLink>. More recently, anti-p53 antibodies are of clinical significance as a serological marker in the diagnosis and monitoring of breast cancer <TextLink reference="32"></TextLink>.</Pgraph>
<Pgraph>To our knowledge, this is the first report to demonstrate the genetic alterations in Egyptian breast cancer patients using FISH technique.</Pgraph>
<Pgraph>In the current study, we assessed the genetic alterations in HER-2/neu, c-myc and p53 genes using fluorescence in situ hybridization (FISH) in Egyptian breast cancer patients and investigated the prognostic role of the three markers and their relation to each other and to demonstrate their relation to the classical clinicopathological factors.</Pgraph>
</TextBlock>
<TextBlock linked="yes" name="Materials and methods">
<MainHeadline>Materials and methods</MainHeadline>
<SubHeadline>Patients</SubHeadline>
<Pgraph>Patients with primary breast cancer had either an excision biopsy with axillary evacuation or a modified radical mastectomy in the surgical department at the National Cancer Institute, Cairo University. No preoperative radiation therapy or chemotherapy was administered to patients.</Pgraph>
<Pgraph>Thirty-four consecutive samples were obtained during the period from 1999–2001 from 33 females and 1 male with breast carcinoma (27 with invasive duct carcinoma and 7 with non-invasive duct carcinoma). The age of the patients ranged from 23 to 75 years. Seventeen samples were obtained from 16 females and 1 male with benign breast lesions to serve as a control group (12 with fibroadenoma, 4 with fibrocyctic disease and 1 with gynecomastia). The age of the patients ranged from 15–52 years. The histologic grades of the tumor foci were assessed using the modified Bloom-Richardson classification <TextLink reference="16"></TextLink>. Clinical staging was done according to TNM classification <TextLink reference="3"></TextLink>. Informed consent was obtained for all cases.</Pgraph>
<Pgraph>Tissue samples were cut into pieces, one was formaline-fixed and paraffin-embedded for subsequent histopathological examination, and the second was shock-freezed in liquid nitrogen and stored at –80°C for the analysis of HER-2/neu, c-myc and p53.</Pgraph>
<SubHeadline>Fluorescence in situ hybridization analysis</SubHeadline>
<Pgraph>Specimen preparation, hybridization, and microscopy were performed as previously described <TextLink reference="37"></TextLink>, <TextLink reference="38"></TextLink> with the following modifications. Frozen-stored tissues were slowly thawed in cold phosphate buffered saline (PBS) at 4°C, light-touched on glass slides. Slides were incubated in 75 mM KCl for 20 min at 37°C, fixed in freshly prepared Carnoy’s solution (methanol : acetic acid, 3:1 v/v ) and air-dried. </Pgraph>
<Pgraph>The SpectrumOrange labeled HER-2/neu probe and the SpectrumGreen chromosome 17 centromeric alpha-satellite (chr.17cen.) probe were cohybridized on the tissue specimen. For c-myc hybridization locus specific identifier (LSI) c-myc (SpectrumOrange) and CEP8 (SpectrumGreen) probe and LSI p53 probe were applied. FISH probes were obtained from Vysis, Inc, Downers Grove, IL. </Pgraph>
<Pgraph>The c-myc and HER-2 gene copy numbers in approximately 200 nuclei in the predominant tumor cell population were estimated in relation with centromere CEP8 and CEP17. Hybridization signals were enumerated by the ratio of orange signals (for c-myc and HER-2) to green signals (for CEP8 and CEP17) in morphologically intact and nonoverlapping nuclei. At least a 3-fold increase of the c-myc signals over CEP8 signals in the tumor cells was considered the criterion for gene amplification. The same criterion was applied to HER-2 amplification analysis using LSI HER-2 probe together with CEP17 probe <TextLink reference="24"></TextLink>, <TextLink reference="9"></TextLink>.</Pgraph>
<SubHeadline>Statistical analysis</SubHeadline>
<Pgraph>Cut-off values were determined using receiver operating characteristic curve (ROC curve). The Mann-Whitney and Wilcoxon tests, and univariate analysis were performed using a chi-square test of association or Fisher’s exact model to test the association of categorical variables, whereas t-test was used for continuous variables. P values below 0.05 were considered as significant <TextLink reference="2"></TextLink>. </Pgraph>
</TextBlock>
<TextBlock linked="yes" name="Results">
<MainHeadline>Results</MainHeadline>
<SubHeadline>Cut-off values for HER-2/neu and c-myc</SubHeadline>
<Pgraph>The best cut-off values were estimated to maximize the sum of sensitivity and specificity. Optimal cut-off value of 1.2 for both HER-2/neu and c-myc are shown in <TextGroup><PlainText>Figure 1 </PlainText></TextGroup><ImgLink imgNo="1" imgType="figure"/>.</Pgraph>
<SubHeadline>Expression of HER-2/neu, c-myc and p53 in benign and malignant groups</SubHeadline>
<Pgraph>Genetic alteration in HER-2/neu, c-myc and p53 were significantly higher in the malignant group when compared to the benign one as show in Table 1 <ImgLink imgNo="1" imgType="table"/>. In the malignant group, the levels (mean, median and positivity rates) for HER-2/neu are 2.8±0.17, 1.435 (0.9–4.2) and 85% respectively, versus 1.25±0.11, 1.06 (0.86–2.5) and 29% in the benign group.</Pgraph>
<Pgraph>For c-myc, the levels (mean, median and positivity rates) are 1.89±0.13, 1,825 (0.95–3.53) and 73% respectively in the malignant group as compared to 1.2±0.08, 1.1 (0.87–2.02) and 35% respectively in the benign group. 0 (0–1) and 64% are the median and positivity rate, respectively for p53 in the malignant group in comparison to 1 (0–1) and 17% in the benign group.</Pgraph>
<SubHeadline>Relation between the investigated molecular markers and clinicopathological findings</SubHeadline>
<Pgraph>The levels of the three investigated genes in relation to the clinicopathological factors were shown in Table 2 <ImgLink imgNo="2" imgType="table"/> and Table 3 <ImgLink imgNo="3" imgType="table"/>. The results show positivity rate of HER-2/neu, c-myc and p53 and their relation to clinicopathological factors. At age <45, positivity rate of HER-2/neu, c-myc and p53 were 12 (75%), 10 (62.5%) and 4 (25%) respectively, whereas at age >45 positivity rate of HER-2/neu c-myc and p53 were 17 (94.44%), 15 (83.33%) and 18 (100%) respectively.</Pgraph>
<Pgraph>In relation to tumor size, the positivity rate of HER-2/neu, c-myc and p53 were 7 (70%), 4 (40%) and 5 (50%) for tumor size less than 3 cm respectively. For the tumor size more and equal to 3 cm, the positivity rate of HER-2/neu, c-myc and p53 were 22 (91.66%), 21 (87.5%) and 17 (70.83%) respectively. On the other hand the positivity rate of HER-2/neu, c-myc and p53 were 25 (89.28%), 19 (67.85%) and 19 (67.85%) respectively in grade I & II cases, whereas it was 4 (75%), 6 (100%) and 3 (50%) respectively in grade III. Concerning disease staging, HER-2/neu and c-myc were the only markers that showed significant expression at late stages of disease (HER-2/neu: 2.86 (1.27–4.2), 2.63±0.23; c-myc: 88.88%) than in early stages (HER-2/neu: 1.35 (1.22–3.24), 1.69±0.2; c-myc: 56.25%).</Pgraph>
<Pgraph>Regarding nodal status, p53 and HER-2/neu were s<TextGroup><PlainText>ignif</PlainText></TextGroup>icantly expressed in patients with positive lymph nodes (p53: 77.3%, HER-2/neu: 2.49±0.22) when compared to those with negative lymph nodal status (p 53: 41.7%, HER-2/neu: 1.69±0.25).</Pgraph>
<SubHeadline>Correlation tests</SubHeadline>
<Pgraph>On relating the level of the three biomarkers to each other, they exhibited a significant correlation with each other (HER-2/neu and c-myc r=0.511, p=0.002; p53 and c-myc r=0.356, p=0.03; p53 and HER-2/neu r=0.432, p=0.01) (Table 4) <ImgLink imgNo="4" imgType="table"/>.</Pgraph>
<SubHeadline>Positivity of the molecular markers in benign and malignant groups</SubHeadline>
<Pgraph>The number of cases that give positive results for the three markers together (HER-2/neu, c-myc and p53) constitute 46% and 12% in malignant and benign cases, respectively. On the other hand, the percentage of negativity for the three markers is 3% and 52% in the malignant and benign cases, respectively.</Pgraph>
</TextBlock>
<TextBlock linked="yes" name="Discussion">
<MainHeadline>Discussion</MainHeadline>
<Pgraph>Breast cancer is a disease with a tremendous heterogeneity in its clinical behavior. It is well known that the clinical outcome of breast cancer patients depends on different clinicopathological factors, including the metastatic status of the lymph nodes, tumor size, tumor grade, and histopathological subtype, but there are other molecular markers that can aid the selection of patients for adjuvant therapies. These molecular markers split the apparently uniform population into clear subpopulations so that the therapies offered to patients may become a reflection of their individual therapeutic needs, rather than the assumption that all patients behave as the population median <TextLink reference="22"></TextLink>. </Pgraph>
<Pgraph>In our study, we assessed the genetic alterations in HER-2/neu, c-myc and p53 genes that are commonly reported in human breast carcinomas. In the present study, the gene alterations were studied using FISH technique. FISH technique is a useful method to evaluate metaphase chromosomes or interphase nuclei to detect genetic aberrations in a variety of solid tumor. FISH analysis of interphase nuclei detects genetic aberrations that are difficult to identify by conventional cytogenetic analysis <TextLink reference="55"></TextLink>, <TextLink reference="7"></TextLink>.</Pgraph>
<Pgraph>Our results revealed that HER-2/neu was amplified in 85% of malignant breast tumors, but only in 29% of benign tumors. Similar results were obtained by Grushko et al. <TextLink reference="21"></TextLink> and Park et al. <TextLink reference="35"></TextLink>. In addition, to evaluate the HER-2/neu status at the mRNA and DNA level of breast carcinomas, Vanden Bempt et al. <TextLink reference="51"></TextLink> studied 32 invasive duct carcinomas for the detection of HER-2/neu mRNA levels by RT-PCR. Corresponding paraffin sections were examined by FISH. Only 28 cases showed increased levels of HER-2/neu mRNA. FISH analysis showed corresponding gene amplification in all 28 cases with 2 cases showing peculiar amplification pattern. In almost all cases, amplification of the HER-2/neu gene results in protein overexpression and poor prognosis. Patients whose tumors have HER-2/neu gene amplification have a shorter disease-free survival time than patients whose tumors exhibit a normal HER-2/neu gene copy number <TextLink reference="20"></TextLink>. In addition, Yassin et al. <TextLink reference="57"></TextLink> studied the expression of HER family in Egyptian breast cancer patients and reported that HER-2 expression was encountered in 79.5% by immunohistochemistry versus 81.6% by RT-PCR. Tubbs et al. <TextLink reference="50"></TextLink> using primary FISH testing in 742 consecutive cases of breast cancer, reported 80% of the cases not amplified for HER-2 (HER-2/CEP17=0.8–1.9), whereas 19% (142/742) of cases were HER-2 amplified (HER-2/CEP17>or=2.0).</Pgraph>
<Pgraph>As shown in our results, HER-2/neu positivity significantly associates with tumor size and grade. Similar results were obtained by Almasri and Al Hamad <TextLink reference="1"></TextLink> and Prati et al. <TextLink reference="42"></TextLink>. Regarding nodal metastasis, our results are in agreement with other studies which reported that HER-2/neu amplification has been strongly associated with positive lymph nodes <TextLink reference="44"></TextLink>, <TextLink reference="29"></TextLink>. They suggested that HER-2/neu oncogene may be a marker for aggressive clinical behavior in breast cancer patients. In relation to stage, HER-2/neu amplification was shown to be significantly associated with late stages of the disease, confirming previous reports <TextLink reference="43"></TextLink>, <TextLink reference="47"></TextLink> that detected increased frequency of HER-2/neu amplification, by FISH analysis, in patients suffering from stages later than IIB based on TNM staging system. HER-2/neu amplification detected by FISH also predicted disease-related death independent of classical clinicopathological factors in breast cancer patients who received adjuvant therapy <TextLink reference="15"></TextLink>. </Pgraph>
<Pgraph>Matrix metalloproteinases (MMPs), in particular the gelatinases MMP2 and MMP9, are important mediators of tumour invasion and metastasis. Decock et al. <TextLink reference="14"></TextLink> demonstrated that patients with HER-2/neu overexpressing tumours showed an increase of 27% in plasma MMP2 activity, but not in MMP9, compared with patients without overexpression, suggesting a role for HER-2/neu in the signalling pathways of MMP2 activation in carcinogenesis.</Pgraph>
<Pgraph>In our study, results showed that c-myc was amplified in 73% of malignant breast tumors, but only in 35% of benign lesions. Similarly, Blancato et al. <TextLink reference="11"></TextLink> reported a high proportion (70%) of breast carcinoma were amplified for the c-myc gene. Also, Park et al. <TextLink reference="35"></TextLink> found that 15.2% of the breast cancer specimens were c-myc positive. Blancato et al. <TextLink reference="11"></TextLink> reported that c-myc gene amplification using FISH was correlated with the percentage of tumor cells which expressed high levels of its protein, as det<TextGroup><PlainText>ecte</PlainText></TextGroup>d by immunohistochemistry in invasive cells.</Pgraph>
<Pgraph>Results of the current study demonstrated a significant relation between c-myc positivity and tumor size, grade and stage of the disease. Regarding tumor size, Aulmann et al. <TextLink reference="5"></TextLink> examined c-myc oncogene amplification using FISH in a series of 96 pure ductal carcinoma in situ (DCIS) and observed that c-myc oncogene amplification was significantly associated with larger tumor size. They concluded that c-myc oncogene appears to be involved in the development of a more aggressive phenotype of DCIS. Shanmugham et al. <TextLink reference="46"></TextLink> reported that c-myc expression, assessed by immunohistochemical staining, showed a positive association with increasing grade. With regard to disease staging, Yang et al. <TextLink reference="56"></TextLink> and Aulmann et al. <TextLink reference="4"></TextLink> showed a significant association between c-myc expression and amplification, respectively, and advanced clinical stage of the disease. They implicated the role of c-myc in tumor development and consequently disease progression.</Pgraph>
<Pgraph>Results of the present study revealed that p53 mutations, assessed by FISH, were detected in 64% of malignant cases but only in 17% of benign tumors. This finding is in agreement with the study of Lukas et al. <TextLink reference="28"></TextLink>, which investigated p53 mutation in breast carcinoma in situ (CIS) using immunohistochemical method. They have shown that p53 protein overexpression was identified in 22% of pure intraductal breast carcinomas and in 35% of breast CIS with invasive disease. Recently, Peng et al. <TextLink reference="39"></TextLink> reported an increase in the level of p53 gene expression in breast cancer using molecular beacon imaging technology. In our study, genetic alterations in p53 were detected in benign breast tumors. This is consistent with the study of Rohan et al. <TextLink reference="45"></TextLink> who demonstrated that accumulation of p53 protein in benign disease was associated with an increased risk of progression to breast cancer.</Pgraph>
<Pgraph>As shown in our results, p53 positivity is significantly associated with age of the patient, size and histological grade of the tumor and positivity of lymph nodes. P<TextGroup><PlainText>ietiläine</PlainText></TextGroup>n et al. <TextLink reference="40"></TextLink> analyzed female breast carcinomas immunohistochemically for p53 expression. They found that high fraction of p53-positive nuclei was significantly related to patient age under 70 years. Moreover, Gion et al. <TextLink reference="18"></TextLink> reported that younger age was related to low p53 values in breast cancer patients. Regarding tumor size, Overgaard et al. <TextLink reference="34"></TextLink> demonstrated that TP 53 gene mutations were significantly more frequent in tumors of large size. In addition, Montero et al. <TextLink reference="31"></TextLink> found a significant relation between p53 positivity and tumor size. With regard to the histological grade, our results showed that p53 mutation was significantly associated with low histological grade, a finding that is contradictory to that of Bertheau et al. <TextLink reference="10"></TextLink> and Goel et al. <TextLink reference="19"></TextLink>. They reported that p53 positivity is significantly associated with high histological grade. This controversy could be referred to the small number of grade III tumors included in our study. Regarding lymph node status, Pratap and Shousha <TextLink reference="41"></TextLink> demonstrated that p53 positivity was significantly related to the presence of extensive axillary lymph node metastasis, a finding that is in harmony with our results. In addition, Cianga et al. <TextLink reference="13"></TextLink> reported that 77.2% of the primary breast carcinomas and 75% of the lymph nodes metastases are positive for von Willebrand factor, a marker of the endothelial cells, and this positivity is s<TextGroup><PlainText>ignif</PlainText></TextGroup>icantly correlated with the expression of p53, supporting the idea that angiogenesis is a marker for tumor aggressiveness and p53 could be involved in this process.</Pgraph>
<Pgraph>In conclusion, future studies must be completed that include larger databases of patients, which integrate clinical, pathologic, and molecular oncogene parameters. The merging of these data may provide the clinician with an enhancement of prognostic information that accurately predicts the aggressive phenotype for breast cancer. Thereafter, decisions regarding the necessity to administer and delete adjuvant therapies will gain scientific credibility.</Pgraph>
</TextBlock>
<TextBlock linked="yes" name="Notes">
<MainHeadline>Notes</MainHeadline>
<SubHeadline>Conflicts of interest</SubHeadline>
<Pgraph>None declared</Pgraph>
</TextBlock>
<References linked="yes">
<Reference refNo="1">
<RefAuthor>Almasri NM</RefAuthor>
<RefAuthor>Al Hamad M</RefAuthor>
<RefTitle>Immunohistochemical evaluation of human epidermal growth factor receptor 2 and estrogen and progesterone receptors in breast carcinoma in Jordan</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Breast Cancer Res</RefJournal>
<RefPage>R598-604</RefPage>
<RefTotal>Almasri NM, Al Hamad M. Immunohistochemical evaluation of human epidermal growth factor receptor 2 and estrogen and progesterone receptors in breast carcinoma in Jordan. Breast Cancer Res. 2005;7(5):R598-604. DOI: 10.1186/bcr1200</RefTotal>
<RefLink>http://dx.doi.org/10.1186/bcr1200</RefLink>
</Reference>
<Reference refNo="3">
<RefAuthor>American Joint Committee on Cancer (AJCC)</RefAuthor>
<RefTitle></RefTitle>
<RefYear>1992</RefYear>
<RefBookTitle>Manual for staging of cancer</RefBookTitle>
<RefPage></RefPage>
<RefTotal>American Joint Committee on Cancer (AJCC). Manual for staging of cancer. 4th ed. Philadelpha: Lippincott; 1992.</RefTotal>
</Reference>
<Reference refNo="4">
<RefAuthor>Aulmann S</RefAuthor>
<RefAuthor>Adler N</RefAuthor>
<RefAuthor>Rom J</RefAuthor>
<RefAuthor>Helmchen B</RefAuthor>
<RefAuthor>Schirmacher P</RefAuthor>
<RefAuthor>Sinn HP</RefAuthor>
<RefTitle>c-myc amplifications in primary breast carcinomas and their local recurrences</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>J Clin Pathol</RefJournal>
<RefPage>424-8</RefPage>
<RefTotal>Aulmann S, Adler N, Rom J, Helmchen B, Schirmacher P, Sinn HP. c-myc amplifications in primary breast carcinomas and their local recurrences. J Clin Pathol. 2006;59(4):424-8. DOI: 10.1136/jcp.2005.029264</RefTotal>
<RefLink>http://dx.doi.org/10.1136/jcp.2005.029264</RefLink>
</Reference>
<Reference refNo="5">
<RefAuthor>Aulmann S</RefAuthor>
<RefAuthor>Bentz M</RefAuthor>
<RefAuthor>Sinn HP</RefAuthor>
<RefTitle>C-myc oncogene amplification in ductal carcinoma in situ of the breast</RefTitle>
<RefYear>2002</RefYear>
<RefJournal>Breast Cancer Res Treat</RefJournal>
<RefPage>25-31</RefPage>
<RefTotal>Aulmann S, Bentz M, Sinn HP. C-myc oncogene amplification in ductal carcinoma in situ of the breast. Breast Cancer Res Treat. 2002;74(1):25-31. DOI: 10.1023/A:1016061327812</RefTotal>
<RefLink>http://dx.doi.org/10.1023/A:1016061327812</RefLink>
</Reference>
<Reference refNo="6">
<RefAuthor>Barsyte-Lovejoy D</RefAuthor>
<RefAuthor>Lau SK</RefAuthor>
<RefAuthor>Boutros PC</RefAuthor>
<RefAuthor>Khosravi F</RefAuthor>
<RefAuthor>Jurisica I</RefAuthor>
<RefAuthor>Andrulis IL</RefAuthor>
<RefAuthor>Tsao MS</RefAuthor>
<RefAuthor>Penn LZ</RefAuthor>
<RefTitle>The c-Myc oncogene directly induces the H19 noncoding RNA by allele-specific binding to potentiate tumorigenesis</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>Cancer Res</RefJournal>
<RefPage>5330-7</RefPage>
<RefTotal>Barsyte-Lovejoy D, Lau SK, Boutros PC, Khosravi F, Jurisica I, Andrulis IL, Tsao MS, Penn LZ. The c-Myc oncogene directly induces the H19 noncoding RNA by allele-specific binding to potentiate tumorigenesis. Cancer Res. 2006;66(10):5330-7. DOI: 10.1158/0008-5472.CAN-06-0037</RefTotal>
<RefLink>http://dx.doi.org/10.1158/0008-5472.CAN-06-0037</RefLink>
</Reference>
<Reference refNo="7">
<RefAuthor>Beatty BG</RefAuthor>
<RefAuthor>Bryant R</RefAuthor>
<RefAuthor>Wang W</RefAuthor>
<RefAuthor>Ashikaga T</RefAuthor>
<RefAuthor>Gibson PC</RefAuthor>
<RefAuthor>Leiman G</RefAuthor>
<RefAuthor>Weaver DL</RefAuthor>
<RefTitle>HER-2/neu detection in fine-needle aspirates of breast cancer: fluorescence in situ hybridization and immunocytochemical analysis</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>Am J Clin Pathol</RefJournal>
<RefPage>246-55</RefPage>
<RefTotal>Beatty BG, Bryant R, Wang W, Ashikaga T, Gibson PC, Leiman G, Weaver DL. HER-2/neu detection in fine-needle aspirates of breast cancer: fluorescence in situ hybridization and immunocytochemical analysis. Am J Clin Pathol. 2004;122(2):246-55. DOI: 10.1309/X8UP920UF4XM1C5C</RefTotal>
<RefLink>http://dx.doi.org/10.1309/X8UP920UF4XM1C5C</RefLink>
</Reference>
<Reference refNo="9">
<RefAuthor>Bernardino J</RefAuthor>
<RefAuthor>Apiou F</RefAuthor>
<RefAuthor>Gerbault-Seureau M</RefAuthor>
<RefAuthor>Malfoy B</RefAuthor>
<RefAuthor>Dutrillaux B</RefAuthor>
<RefTitle>Characterization of recurrent homogeneously staining regions in 72 breast carcinomas</RefTitle>
<RefYear>1998</RefYear>
<RefJournal>Genes Chromosomes Cancer</RefJournal>
<RefPage>100-8</RefPage>
<RefTotal>Bernardino J, Apiou F, Gerbault-Seureau M, Malfoy B, Dutrillaux B. Characterization of recurrent homogeneously staining regions in 72 breast carcinomas. Genes Chromosomes Cancer. 1998;23: 100-8. DOI: 10.1002/(SICI)1098-2264(199810)23:2<100::AID-GCC2>3.0.CO;2-6</RefTotal>
<RefLink>http://dx.doi.org/10.1002/(SICI)1098-2264(199810)23:2<100::AID-GCC2>3.0.CO;2-6</RefLink>
</Reference>
<Reference refNo="49">
<RefAuthor>Stewart BW</RefAuthor>
<RefAuthor>Kleihues P</RefAuthor>
<RefTitle></RefTitle>
<RefYear>2003</RefYear>
<RefBookTitle>World Cancer Report</RefBookTitle>
<RefPage></RefPage>
<RefTotal>Stewart BW, Kleihues P, editors. World Cancer Report. Lyon (France): International Agency for Research on Cancer; 2003.</RefTotal>
</Reference>
<Reference refNo="10">
<RefAuthor>Bertheau P</RefAuthor>
<RefAuthor>Steinberg SM</RefAuthor>
<RefAuthor>Merino MJ</RefAuthor>
<RefTitle>C-erbB-2, p53, and nm23 gene product expression in breast cancer in young women: immunohistochemical analysis and clinicopathologic correlation</RefTitle>
<RefYear>1998</RefYear>
<RefJournal>Hum Pathol</RefJournal>
<RefPage>323-9</RefPage>
<RefTotal>Bertheau P, Steinberg SM, Merino MJ. C-erbB-2, p53, and nm23 gene product expression in breast cancer in young women: immunohistochemical analysis and clinicopathologic correlation. Hum Pathol. 1998;29(4):323-9. DOI: 10.1016/S0046-8177(98)90111-3</RefTotal>
<RefLink>http://dx.doi.org/10.1016/S0046-8177(98)90111-3</RefLink>
</Reference>
<Reference refNo="11">
<RefAuthor>Blancato J</RefAuthor>
<RefAuthor>Singh B</RefAuthor>
<RefAuthor>Liu A</RefAuthor>
<RefAuthor>Liao DJ</RefAuthor>
<RefAuthor>Dickson RB</RefAuthor>
<RefTitle>Correlation of amplification and overexpression of the c-myc oncogene in high-grade breast cancer: FISH, in situ hybridisation and immunohistochemical analyses</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>Br J Cancer</RefJournal>
<RefPage>1612-9</RefPage>
<RefTotal>Blancato J, Singh B, Liu A, Liao DJ, Dickson RB. Correlation of amplification and overexpression of the c-myc oncogene in high-grade breast cancer: FISH, in situ hybridisation and immunohistochemical analyses. Br J Cancer. 2004;90(8):1612-9. DOI: 10.1038/sj.bjc.6601703</RefTotal>
<RefLink>http://dx.doi.org/10.1038/sj.bjc.6601703</RefLink>
</Reference>
<Reference refNo="12">
<RefAuthor>Brennan PJ</RefAuthor>
<RefAuthor>Kumagai T</RefAuthor>
<RefAuthor>Berezov A</RefAuthor>
<RefAuthor>Murali R</RefAuthor>
<RefAuthor>Greene MI</RefAuthor>
<RefTitle>HER2/neu: mechanisms of dimerization/oligomerization</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Oncogene</RefJournal>
<RefPage>6093-101</RefPage>
<RefTotal>Brennan PJ, Kumagai T, Berezov A, Murali R, Greene MI. HER2/neu: mechanisms of dimerization/oligomerization. Oncogene. 2000;19:6093-101. DOI: 10.1038/sj.onc.1203967</RefTotal>
<RefLink>http://dx.doi.org/10.1038/sj.onc.1203967</RefLink>
</Reference>
<Reference refNo="13">
<RefAuthor>Cianga C</RefAuthor>
<RefAuthor>Cianga P</RefAuthor>
<RefAuthor>Cozma L</RefAuthor>
<RefAuthor>Diaconu C</RefAuthor>
<RefAuthor>Carasevici E</RefAuthor>
<RefTitle>Evaluarea angiogenezei in cancerul mamar. Corelatii cu agresivitatea tumorala [Evaluation of the angiogenic process in breast carcinoma--correlations with cancer aggressiveness]</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>Rev Med Chir Soc Med Nat Iasi</RefJournal>
<RefPage>657-61</RefPage>
<RefTotal>Cianga C, Cianga P, Cozma L, Diaconu C, Carasevici E. Evaluarea angiogenezei in cancerul mamar. Corelatii cu agresivitatea tumorala [Evaluation of the angiogenic process in breast carcinoma--correlations with cancer aggressiveness]. Rev Med Chir Soc Med Nat Iasi. 2004;108(3):657-61.</RefTotal>
</Reference>
<Reference refNo="14">
<RefAuthor>Decock J</RefAuthor>
<RefAuthor>Hendrickx W</RefAuthor>
<RefAuthor>Wildiers H</RefAuthor>
<RefAuthor>Christiaens MR</RefAuthor>
<RefAuthor>Neven P</RefAuthor>
<RefAuthor>Drijkoningen M</RefAuthor>
<RefAuthor>Paridaens R</RefAuthor>
<RefTitle>Plasma gelatinase levels in patients with primary breast cancer in relation to axillary lymph node status, Her2/neu expression and other clinicopathological variables</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Clin Exp Metastasis</RefJournal>
<RefPage>495-502</RefPage>
<RefTotal>Decock J, Hendrickx W, Wildiers H, Christiaens MR, Neven P, Drijkoningen M, Paridaens R. Plasma gelatinase levels in patients with primary breast cancer in relation to axillary lymph node status, Her2/neu expression and other clinicopathological variables. Clin Exp Metastasis. 2005;22(6):495-502. DOI: 10.1007/s10585-005-3992-2</RefTotal>
<RefLink>http://dx.doi.org/10.1007/s10585-005-3992-2</RefLink>
</Reference>
<Reference refNo="15">
<RefAuthor>Depowski PL</RefAuthor>
<RefAuthor>Brien TP</RefAuthor>
<RefAuthor>Sheehan CE</RefAuthor>
<RefAuthor>Stylos S</RefAuthor>
<RefAuthor>Johnson RL</RefAuthor>
<RefAuthor>Ross JS</RefAuthor>
<RefTitle>Prognostic significance of p34cdc2 cyclin-dependent kinase and MIB1 overexpression, and HER-2/neu gene amplification detected by fluorescence in situ hybridization in breast cancer</RefTitle>
<RefYear>1999</RefYear>
<RefJournal>Am J Clin Pathol</RefJournal>
<RefPage>459-69</RefPage>
<RefTotal>Depowski PL, Brien TP, Sheehan CE, Stylos S, Johnson RL, Ross JS. Prognostic significance of p34cdc2 cyclin-dependent kinase and MIB1 overexpression, and HER-2/neu gene amplification detected by fluorescence in situ hybridization in breast cancer. Am J Clin Pathol. 1999;112(4):459-69.</RefTotal>
</Reference>
<Reference refNo="16">
<RefAuthor>Elston CW</RefAuthor>
<RefAuthor>Ellis IO</RefAuthor>
<RefTitle>Pathological prognostic factors in breast cancer: the value of histologic grade in breast cancer: Experience from a large study with long term follow-up</RefTitle>
<RefYear>1991</RefYear>
<RefJournal>Histopathology</RefJournal>
<RefPage>403-10</RefPage>
<RefTotal>Elston CW, Ellis IO. Pathological prognostic factors in breast cancer: the value of histologic grade in breast cancer: Experience from a large study with long term follow-up. Histopathology. 1991;19:403-10. DOI: 10.1111/j.1365-2559.1991.tb00229.x</RefTotal>
<RefLink>http://dx.doi.org/10.1111/j.1365-2559.1991.tb00229.x</RefLink>
</Reference>
<Reference refNo="17">
<RefAuthor>Ferlay J</RefAuthor>
<RefAuthor>Bray F</RefAuthor>
<RefAuthor>Pisani P</RefAuthor>
<RefAuthor>Parkin DM</RefAuthor>
<RefTitle></RefTitle>
<RefYear>2004</RefYear>
<RefBookTitle>GLOBOCAN 2002: Cancer Incidence, Mortality and Prevalence Worldwide IARC CancerBase No 5, version 2.0</RefBookTitle>
<RefPage></RefPage>
<RefTotal>Ferlay J, Bray F, Pisani P, Parkin DM. GLOBOCAN 2002: Cancer Incidence, Mortality and Prevalence Worldwide IARC CancerBase No 5, version 2.0. Lyon, France: IARC Press; 2004.</RefTotal>
</Reference>
<Reference refNo="18">
<RefAuthor>Gion M</RefAuthor>
<RefAuthor>Boracchi P</RefAuthor>
<RefAuthor>Dittadi R</RefAuthor>
<RefAuthor>Biganzoli E</RefAuthor>
<RefAuthor>Peloso L</RefAuthor>
<RefAuthor>Gatti C</RefAuthor>
<RefAuthor>Paccagnella A</RefAuthor>
<RefAuthor>Rosabian A</RefAuthor>
<RefAuthor>Vinante O</RefAuthor>
<RefAuthor>Meo S</RefAuthor>
<RefTitle>Quantitative measurement of soluble cytokeratin fragments in tissue cytosol of 599 node negative breast cancer patients: a prognostic marker possibly associated with apoptosis</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Breast Cancer Res Treat</RefJournal>
<RefPage>211-21</RefPage>
<RefTotal>Gion M, Boracchi P, Dittadi R, Biganzoli E, Peloso L, Gatti C, Paccagnella A, Rosabian A, Vinante O, Meo S. Quantitative measurement of soluble cytokeratin fragments in tissue cytosol of 599 node negative breast cancer patients: a prognostic marker possibly associated with apoptosis. Breast Cancer Res Treat. 2000;59(3):211-21. DOI: 10.1023/A:1006318112776</RefTotal>
<RefLink>http://dx.doi.org/10.1023/A:1006318112776</RefLink>
</Reference>
<Reference refNo="19">
<RefAuthor>Goel MM</RefAuthor>
<RefAuthor>Goel R</RefAuthor>
<RefAuthor>Mehrotra A</RefAuthor>
<RefAuthor>Nath P</RefAuthor>
<RefAuthor>Agarwal PK</RefAuthor>
<RefAuthor>Singh K</RefAuthor>
<RefAuthor>Mehrotra R</RefAuthor>
<RefTitle>Immunohistochemical localization and correlation of p53 and PCNA expression in breast carcinoma</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Indian J Exp Biol</RefJournal>
<RefPage>225-30</RefPage>
<RefTotal>Goel MM, Goel R, Mehrotra A, Nath P, Agarwal PK, Singh K, Mehrotra R. Immunohistochemical localization and correlation of p53 and PCNA expression in breast carcinoma. Indian J Exp Biol. 2000;38(3):225-30.</RefTotal>
</Reference>
<Reference refNo="20">
<RefAuthor>Gonzalez-Angulo AM</RefAuthor>
<RefAuthor>Hortobagyi GN</RefAuthor>
<RefAuthor>Esteva FJ</RefAuthor>
<RefTitle>Adjuvant therapy with trastuzumab for HER-2/neu-positive breast cancer</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>Oncologist</RefJournal>
<RefPage>857-67</RefPage>
<RefTotal>Gonzalez-Angulo AM, Hortobagyi GN, Esteva FJ. Adjuvant therapy with trastuzumab for HER-2/neu-positive breast cancer. Oncologist. 2006;11(8):857-67. DOI: 10.1634/theoncologist.11-8-857</RefTotal>
<RefLink>http://dx.doi.org/10.1634/theoncologist.11-8-857</RefLink>
</Reference>
<Reference refNo="21">
<RefAuthor>Grushko TA</RefAuthor>
<RefAuthor>Blackwood MA</RefAuthor>
<RefAuthor>Schumm PL</RefAuthor>
<RefAuthor>Hagos FG</RefAuthor>
<RefAuthor>Adeyanju MO</RefAuthor>
<RefAuthor>Feldman MD</RefAuthor>
<RefAuthor>Sanders MO</RefAuthor>
<RefAuthor>Weber BL</RefAuthor>
<RefAuthor>Olopade OI</RefAuthor>
<RefTitle>Molecular-cytogenetic analysis of HER-2/neu gene in BRCA1-associated breast cancers</RefTitle>
<RefYear>2002</RefYear>
<RefJournal>Cancer Res</RefJournal>
<RefPage>1481-8</RefPage>
<RefTotal>Grushko TA, Blackwood MA, Schumm PL, Hagos FG, Adeyanju MO, Feldman MD, Sanders MO, Weber BL, Olopade OI. Molecular-cytogenetic analysis of HER-2/neu gene in BRCA1-associated breast cancers. Cancer Res. 2002;62(5):1481-8.</RefTotal>
</Reference>
<Reference refNo="22">
<RefAuthor>Hamilton A</RefAuthor>
<RefAuthor>Piccart M</RefAuthor>
<RefTitle>The contribution of molecular markers to the prediction of response in the treatment of breast cancer: A review of the literature on HER-2, p53 and BCL-2</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Ann Oncol</RefJournal>
<RefPage>647-63</RefPage>
<RefTotal>Hamilton A, Piccart M. The contribution of molecular markers to the prediction of response in the treatment of breast cancer: A review of the literature on HER-2, p53 and BCL-2. Ann Oncol. 2000;11(6):647-63. DOI: 10.1023/A:1008390429428</RefTotal>
<RefLink>http://dx.doi.org/10.1023/A:1008390429428</RefLink>
</Reference>
<Reference refNo="23">
<RefAuthor>Harari D</RefAuthor>
<RefAuthor>Yarden Y</RefAuthor>
<RefTitle>Molecular mechanisms underlying ErbB2/HER2 action in breast cancer</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Oncogene</RefJournal>
<RefPage>6102-14</RefPage>
<RefTotal>Harari D, Yarden Y. Molecular mechanisms underlying ErbB2/HER2 action in breast cancer. Oncogene. 2000;19:6102-14. DOI: 10.1038/sj.onc.1203973</RefTotal>
<RefLink>http://dx.doi.org/10.1038/sj.onc.1203973</RefLink>
</Reference>
<Reference refNo="24">
<RefAuthor>Kallioniemi OP</RefAuthor>
<RefAuthor>Kallioniemi A</RefAuthor>
<RefAuthor>Kurisu W</RefAuthor>
<RefAuthor>Thor A</RefAuthor>
<RefAuthor>Chen LC</RefAuthor>
<RefAuthor>Smith HS</RefAuthor>
<RefAuthor>Waldman FM</RefAuthor>
<RefAuthor>Pinkel D</RefAuthor>
<RefAuthor>Gray JW</RefAuthor>
<RefTitle>ERBB2 amplification in breast cancer analysed by fluorescence in situ hybridization</RefTitle>
<RefYear>1992</RefYear>
<RefJournal>Proc Natl Acad Sci U S A</RefJournal>
<RefPage>5321-5</RefPage>
<RefTotal>Kallioniemi OP, Kallioniemi A, Kurisu W, Thor A, Chen LC, Smith HS, Waldman FM, Pinkel D, Gray JW. ERBB2 amplification in breast cancer analysed by fluorescence in situ hybridization. Proc Natl Acad Sci U S A. 1992;89(12):5321-5. DOI: 10.1073/pnas.89.12.5321</RefTotal>
<RefLink>http://dx.doi.org/10.1073/pnas.89.12.5321</RefLink>
</Reference>
<Reference refNo="25">
<RefAuthor>Lane DP</RefAuthor>
<RefTitle>Cancer: p53, guardian of the genome</RefTitle>
<RefYear>1992</RefYear>
<RefJournal>Nature</RefJournal>
<RefPage>15-6</RefPage>
<RefTotal>Lane DP. Cancer: p53, guardian of the genome. Nature. 1992;358:15-6. DOI: 10.1038/358015a0</RefTotal>
<RefLink>http://dx.doi.org/10.1038/358015a0</RefLink>
</Reference>
<Reference refNo="26">
<RefAuthor>Liao DJ</RefAuthor>
<RefAuthor>Dickson R</RefAuthor>
<RefTitle>c-Myc in breast cancer</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Endocr Relat Cancer</RefJournal>
<RefPage>143-64</RefPage>
<RefTotal>Liao DJ, Dickson R. c-Myc in breast cancer. Endocr Relat Cancer. 2000;7(3):143-64. DOI: 10.1677/erc.0.0070143</RefTotal>
<RefLink>http://dx.doi.org/10.1677/erc.0.0070143</RefLink>
</Reference>
<Reference refNo="27">
<RefAuthor>Lotem J</RefAuthor>
<RefAuthor>Sachs L</RefAuthor>
<RefTitle>Regulation by bcl-2, c-myc and p53 of susceptibility to induction of apoptosis by heat shock and cancer chemotherapy compounds in differentiation-competent and -defective myeloid leukemic cells</RefTitle>
<RefYear>1993</RefYear>
<RefJournal>Cell Growth Differ</RefJournal>
<RefPage>41-7</RefPage>
<RefTotal>Lotem J, Sachs L. Regulation by bcl-2, c-myc and p53 of susceptibility to induction of apoptosis by heat shock and cancer chemotherapy compounds in differentiation-competent and -defective myeloid leukemic cells. Cell Growth Differ. 1993;4(1):41-7.</RefTotal>
</Reference>
<Reference refNo="28">
<RefAuthor>Lukas J</RefAuthor>
<RefAuthor>Niu N</RefAuthor>
<RefAuthor>Press MF</RefAuthor>
<RefTitle>p53 mutations and expression in breast carcinoma in situ</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Am J Pathol</RefJournal>
<RefPage>183-91</RefPage>
<RefTotal>Lukas J, Niu N, Press MF. p53 mutations and expression in breast carcinoma in situ. Am J Pathol. 2000;156(1):183-91.</RefTotal>
</Reference>
<Reference refNo="29">
<RefAuthor>Maru D</RefAuthor>
<RefAuthor>Middleton LP</RefAuthor>
<RefAuthor>Wang S</RefAuthor>
<RefAuthor>Valero V</RefAuthor>
<RefAuthor>Sahin A</RefAuthor>
<RefTitle>HER-2/neu and p53 overexpression as biomarkers of breast carcinoma in women age 30 years and younger</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Cancer</RefJournal>
<RefPage>900-5</RefPage>
<RefTotal>Maru D, Middleton LP, Wang S, Valero V, Sahin A. HER-2/neu and p53 overexpression as biomarkers of breast carcinoma in women age 30 years and younger. Cancer. 2005;103(5): 900-5. DOI: 10.1002/cncr.20850</RefTotal>
<RefLink>http://dx.doi.org/10.1002/cncr.20850</RefLink>
</Reference>
<Reference refNo="30">
<RefAuthor>Miller C</RefAuthor>
<RefAuthor>Mohandas T</RefAuthor>
<RefAuthor>Wolf D</RefAuthor>
<RefAuthor>Prokocimer M</RefAuthor>
<RefAuthor>Rotter V</RefAuthor>
<RefAuthor>Koeffler HP</RefAuthor>
<RefTitle>Human p53 gene localized to short arm of chromosome 17</RefTitle>
<RefYear>1986</RefYear>
<RefJournal>Nature</RefJournal>
<RefPage>783-4</RefPage>
<RefTotal>Miller C, Mohandas T, Wolf D, Prokocimer M, Rotter V, Koeffler HP. Human p53 gene localized to short arm of chromosome 17. Nature. 1986;319(6056):783-4. DOI: 10.1038/319783a0</RefTotal>
<RefLink>http://dx.doi.org/10.1038/319783a0</RefLink>
</Reference>
<Reference refNo="31">
<RefAuthor>Montero S</RefAuthor>
<RefAuthor>Guzmán C</RefAuthor>
<RefAuthor>Vargas C</RefAuthor>
<RefAuthor>Ballestín C</RefAuthor>
<RefAuthor>Cortés-Funes H</RefAuthor>
<RefAuthor>Colomer R</RefAuthor>
<RefTitle>Prognostic value of cytosolic p53 protein in breast cancer</RefTitle>
<RefYear>2001</RefYear>
<RefJournal>Tumor Biol</RefJournal>
<RefPage>337-44</RefPage>
<RefTotal>Montero S, Guzmán C, Vargas C, Ballestín C, Cortés-Funes H, Colomer R. Prognostic value of cytosolic p53 protein in breast cancer. Tumor Biol. 2001;22(5):337-44. DOI: 10.1159/000050636</RefTotal>
<RefLink>http://dx.doi.org/10.1159/000050636</RefLink>
</Reference>
<Reference refNo="32">
<RefAuthor>Müller M</RefAuthor>
<RefAuthor>Meyer M</RefAuthor>
<RefAuthor>Schilling T</RefAuthor>
<RefAuthor>Ulsperger E</RefAuthor>
<RefAuthor>Lehnert T</RefAuthor>
<RefAuthor>Zentgraf H</RefAuthor>
<RefAuthor>Stremmel W</RefAuthor>
<RefAuthor>Volkmann M</RefAuthor>
<RefAuthor>Galle PR</RefAuthor>
<RefTitle>Testing for anti-p53 antibodies increases the diagnostic sensitivity of conventional tumor markers</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>Int J Oncol</RefJournal>
<RefPage>973-80</RefPage>
<RefTotal>Müller M, Meyer M, Schilling T, Ulsperger E, Lehnert T, Zentgraf H, Stremmel W, Volkmann M, Galle PR. Testing for anti-p53 antibodies increases the diagnostic sensitivity of conventional tumor markers. Int J Oncol. 2006;29(4):973-80.</RefTotal>
</Reference>
<Reference refNo="33">
<RefAuthor>Nesbit CE</RefAuthor>
<RefAuthor>Tersak JM</RefAuthor>
<RefAuthor>Prochownick EV</RefAuthor>
<RefTitle>MYC oncogenes and human neoplastic disease</RefTitle>
<RefYear>1999</RefYear>
<RefJournal>Oncogene</RefJournal>
<RefPage>3004-16</RefPage>
<RefTotal>Nesbit CE, Tersak JM, Prochownick EV. MYC oncogenes and human neoplastic disease. Oncogene. 1999;18:3004-16. DOI: 10.1038/sj.onc.1202746</RefTotal>
<RefLink>http://dx.doi.org/10.1038/sj.onc.1202746</RefLink>
</Reference>
<Reference refNo="34">
<RefAuthor>Overgaard J</RefAuthor>
<RefAuthor>Yilmaz M</RefAuthor>
<RefAuthor>Guldberg P</RefAuthor>
<RefAuthor>Hansen LL</RefAuthor>
<RefAuthor>Alsner J</RefAuthor>
<RefTitle>TP53 mutation is an independent prognostic marker for poor outcome in both node-negative and node-positive breast cancer</RefTitle>
<RefYear>2000</RefYear>
<RefJournal>Acta Oncol</RefJournal>
<RefPage>327-33</RefPage>
<RefTotal>Overgaard J, Yilmaz M, Guldberg P, Hansen LL, Alsner J. TP53 mutation is an independent prognostic marker for poor outcome in both node-negative and node-positive breast cancer. Acta Oncol. 2000;39(3):327-33. DOI: 10.1080/028418600750013096</RefTotal>
<RefLink>http://dx.doi.org/10.1080/028418600750013096</RefLink>
</Reference>
<Reference refNo="35">
<RefAuthor>Park K</RefAuthor>
<RefAuthor>Kwak K</RefAuthor>
<RefAuthor>Kim J</RefAuthor>
<RefAuthor>Lim S</RefAuthor>
<RefAuthor>Han S</RefAuthor>
<RefTitle>c-myc amplification is associated with HER2 amplification and closely linked with cell proliferation in tissue microarray of nonselected breast cancers</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Hum Pathol</RefJournal>
<RefPage>634-9</RefPage>
<RefTotal>Park K, Kwak K, Kim J, Lim S, Han S. c-myc amplification is associated with HER2 amplification and closely linked with cell proliferation in tissue microarray of nonselected breast cancers. Hum Pathol. 2005;36(6):634-9. DOI: 10.1016/j.humpath.2005.04.016</RefTotal>
<RefLink>http://dx.doi.org/10.1016/j.humpath.2005.04.016</RefLink>
</Reference>
<Reference refNo="36">
<RefAuthor>Parkin DM</RefAuthor>
<RefAuthor>Whelan SL</RefAuthor>
<RefAuthor>Ferlay J</RefAuthor>
<RefAuthor>Teppo L</RefAuthor>
<RefTitle></RefTitle>
<RefYear>2002</RefYear>
<RefBookTitle>Cancer incidence in five continents, volume VIII</RefBookTitle>
<RefPage></RefPage>
<RefTotal>Parkin DM, Whelan SL, Ferlay J, Teppo L, editors. Cancer incidence in five continents, volume VIII. Lyon (France): International Agency for Research on Cancer; 2002. (IARC Scientific Publication; No. 155).</RefTotal>
</Reference>
<Reference refNo="37">
<RefAuthor>Pauletti G</RefAuthor>
<RefAuthor>Godolphin W</RefAuthor>
<RefAuthor>Press MF</RefAuthor>
<RefAuthor>Slamon DJ</RefAuthor>
<RefTitle>Detection and quantitation of HER-2/neu gene amplification in human breast cancer archival material using fluorescence in situ hybridization</RefTitle>
<RefYear>1996</RefYear>
<RefJournal>Oncogene</RefJournal>
<RefPage>63-72</RefPage>
<RefTotal>Pauletti G, Godolphin W, Press MF, Slamon DJ. Detection and quantitation of HER-2/neu gene amplification in human breast cancer archival material using fluorescence in situ hybridization. Oncogene. 1996;13(1): 63-72.</RefTotal>
</Reference>
<Reference refNo="38">
<RefAuthor>Pauletti G</RefAuthor>
<RefAuthor>Slamon DJ</RefAuthor>
<RefTitle>Interphase cytogenetics of the HER-2/neu oncogene in breast cancer</RefTitle>
<RefYear>1999</RefYear>
<RefBookTitle>Introduction to Fluorescence In Situ Hybridization: Principles and Clinical Application</RefBookTitle>
<RefPage>153-71</RefPage>
<RefTotal>Pauletti G, Slamon DJ. Interphase cytogenetics of the HER-2/neu oncogene in breast cancer. In: Andreeff M, Pinkel D, editors. Introduction to Fluorescence In Situ Hybridization: Principles and Clinical Application. New York, NY: Wiley-Liss; 1999. p. 153-71.</RefTotal>
</Reference>
<Reference refNo="39">
<RefAuthor>Peng XH</RefAuthor>
<RefAuthor>Cao ZH</RefAuthor>
<RefAuthor>Xia JT</RefAuthor>
<RefAuthor>Carlson GW</RefAuthor>
<RefAuthor>Lewis MM</RefAuthor>
<RefAuthor>Wood WC</RefAuthor>
<RefAuthor>Yang L</RefAuthor>
<RefTitle>Real-time detection of gene expression in cancer cells using molecular beacon imaging: new strategies for cancer research</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Cancer Res</RefJournal>
<RefPage>1909-17</RefPage>
<RefTotal>Peng XH, Cao ZH, Xia JT, Carlson GW, Lewis MM, Wood WC, Yang L. Real-time detection of gene expression in cancer cells using molecular beacon imaging: new strategies for cancer research. Cancer Res. 2005;65(5): 1909-17. DOI: 10.1158/0008-5472.CAN-04-3196</RefTotal>
<RefLink>http://dx.doi.org/10.1158/0008-5472.CAN-04-3196</RefLink>
</Reference>
<Reference refNo="40">
<RefAuthor>Pietiläinen T</RefAuthor>
<RefAuthor>Lipponen P</RefAuthor>
<RefAuthor>Aaltomaa S</RefAuthor>
<RefAuthor>Eskelinen M</RefAuthor>
<RefAuthor>Kosma VM</RefAuthor>
<RefAuthor>Syrjänen K</RefAuthor>
<RefTitle>Expression of p53 protein has no independent prognostic value in breast cancer</RefTitle>
<RefYear>1995</RefYear>
<RefJournal>J Pathol</RefJournal>
<RefPage>225-32</RefPage>
<RefTotal>Pietiläinen T, Lipponen P, Aaltomaa S, Eskelinen M, Kosma VM, Syrjänen K. Expression of p53 protein has no independent prognostic value in breast cancer. J Pathol. 1995;177(3):225-32. DOI: 10.1002/path.1711770303</RefTotal>
<RefLink>http://dx.doi.org/10.1002/path.1711770303</RefLink>
</Reference>
<Reference refNo="41">
<RefAuthor>Pratap R</RefAuthor>
<RefAuthor>Shousha S</RefAuthor>
<RefTitle>Breast carcinoma in women under the age of 50: Relationship between p53 immunostaining, tumour grade, and axillary lymph node status</RefTitle>
<RefYear>1998</RefYear>
<RefJournal>Breast Cancer Res Treat</RefJournal>
<RefPage>35-9</RefPage>
<RefTotal>Pratap R, Shousha S. Breast carcinoma in women under the age of 50: Relationship between p53 immunostaining, tumour grade, and axillary lymph node status. Breast Cancer Res Treat. 1998;49(1):35-9. DOI: 10.1023/A:1005993220824</RefTotal>
<RefLink>http://dx.doi.org/10.1023/A:1005993220824</RefLink>
</Reference>
<Reference refNo="42">
<RefAuthor>Prati R</RefAuthor>
<RefAuthor>Apple SK</RefAuthor>
<RefAuthor>He J</RefAuthor>
<RefAuthor>Gornbein JA</RefAuthor>
<RefAuthor>Chang HR</RefAuthor>
<RefTitle>Histopathologic characteristics predicting HER-2/neu amplification in breast cancer</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Breast J</RefJournal>
<RefPage>433-9</RefPage>
<RefTotal>Prati R, Apple SK, He J, Gornbein JA, Chang HR. Histopathologic characteristics predicting HER-2/neu amplification in breast cancer. Breast J. 2005;11(6):433-9. DOI: 10.1111/j.1075-122X.2005.00125.x</RefTotal>
<RefLink>http://dx.doi.org/10.1111/j.1075-122X.2005.00125.x</RefLink>
</Reference>
<Reference refNo="43">
<RefAuthor>Regitnig P</RefAuthor>
<RefAuthor>Schippinger W</RefAuthor>
<RefAuthor>Lindbauer M</RefAuthor>
<RefAuthor>Samonigg H</RefAuthor>
<RefAuthor>Lax SF</RefAuthor>
<RefTitle>Change of HER-2/neu status in a subset of distant metastases from breastcarcinomas</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>J Pathol</RefJournal>
<RefPage>918-26</RefPage>
<RefTotal>Regitnig P, Schippinger W, Lindbauer M, Samonigg H, Lax SF. Change of HER-2/neu status in a subset of distant metastases from breastcarcinomas. J Pathol. 2004;203(4):918-26. DOI: 10.1002/path.1592</RefTotal>
<RefLink>http://dx.doi.org/10.1002/path.1592</RefLink>
</Reference>
<Reference refNo="44">
<RefAuthor>Riou G</RefAuthor>
<RefAuthor>Mathieu MC</RefAuthor>
<RefAuthor>Barrois M</RefAuthor>
<RefAuthor>Le Bihan ML</RefAuthor>
<RefAuthor>Ahomadegbe JC</RefAuthor>
<RefAuthor>Bénard J</RefAuthor>
<RefAuthor>Lê MG</RefAuthor>
<RefTitle>c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein overexpression in operable breast cancer patients</RefTitle>
<RefYear>2001</RefYear>
<RefJournal>Int J Cancer</RefJournal>
<RefPage>266-70</RefPage>
<RefTotal>Riou G, Mathieu MC, Barrois M, Le Bihan ML, Ahomadegbe JC, Bénard J, Lê MG. c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein overexpression in operable breast cancer patients . Int J Cancer. 2001;95(4):266-70.</RefTotal>
</Reference>
<Reference refNo="45">
<RefAuthor>Rohan TE</RefAuthor>
<RefAuthor>Hartwick W</RefAuthor>
<RefAuthor>Miller AB</RefAuthor>
<RefAuthor>Kandel RA</RefAuthor>
<RefTitle>Immunohistochemical detection of c-erbB-2 and p53 in benign breast disease and breast cancer risk</RefTitle>
<RefYear>1998</RefYear>
<RefJournal>J Natl Cancer Inst</RefJournal>
<RefPage>1262-9</RefPage>
<RefTotal>Rohan TE, Hartwick W, Miller AB, Kandel RA. Immunohistochemical detection of c-erbB-2 and p53 in benign breast disease and breast cancer risk. J Natl Cancer Inst. 1998;90(17):1262-9. DOI: 10.1093/jnci/90.17.1262</RefTotal>
<RefLink>http://dx.doi.org/10.1093/jnci/90.17.1262</RefLink>
</Reference>
<Reference refNo="46">
<RefAuthor>Shanmugham R</RefAuthor>
<RefAuthor>Gopalan R</RefAuthor>
<RefAuthor>Shanthi P</RefAuthor>
<RefAuthor>Krishnan KB</RefAuthor>
<RefTitle>Tumor angiogenesis and C-myc expression in breast carcinomas</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>Indian J Pathol Microbiol</RefJournal>
<RefPage>340-2</RefPage>
<RefTotal>Shanmugham R, Gopalan R, Shanthi P, Krishnan KB. Tumor angiogenesis and C-myc expression in breast carcinomas. Indian J Pathol Microbiol. 2004;47(3):340-2.</RefTotal>
</Reference>
<Reference refNo="47">
<RefAuthor>Shimada M</RefAuthor>
<RefAuthor>Imura J</RefAuthor>
<RefAuthor>Kozaki T</RefAuthor>
<RefAuthor>Fujimori T</RefAuthor>
<RefAuthor>Asakawa S</RefAuthor>
<RefAuthor>Shimizu N</RefAuthor>
<RefAuthor>Kawaguchi R</RefAuthor>
<RefTitle>Detection of Her2/neu, c-MYC and ZNF217 gene amplification during breast cancer progression using fluorescence in situ hybridization</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Oncol Rep</RefJournal>
<RefPage>633-41</RefPage>
<RefTotal>Shimada M, Imura J, Kozaki T, Fujimori T, Asakawa S, Shimizu N, Kawaguchi R. Detection of Her2/neu, c-MYC and ZNF217 gene amplification during breast cancer progression using fluorescence in situ hybridization. Oncol Rep. 2005;13(4):633-41.</RefTotal>
</Reference>
<Reference refNo="48">
<RefAuthor>Slamon DJ</RefAuthor>
<RefAuthor>Clark GM</RefAuthor>
<RefAuthor>Wong SG</RefAuthor>
<RefAuthor>Levin WJ</RefAuthor>
<RefAuthor>Ullrich A</RefAuthor>
<RefAuthor>McGuire WL</RefAuthor>
<RefTitle>Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene</RefTitle>
<RefYear>1987</RefYear>
<RefJournal>Science</RefJournal>
<RefPage>177-82</RefPage>
<RefTotal>Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 1987;235(4785):177-82. DOI: 10.1126/science.3798106</RefTotal>
<RefLink>http://dx.doi.org/10.1126/science.3798106</RefLink>
</Reference>
<Reference refNo="8">
<RefAuthor>Benchimol S</RefAuthor>
<RefAuthor>Minden MD</RefAuthor>
<RefTitle>Viruses, Oncogenes and tumor suppressor genes</RefTitle>
<RefYear>1998</RefYear>
<RefBookTitle>The basic science of oncology</RefBookTitle>
<RefPage>79-105</RefPage>
<RefTotal>Benchimol S, Minden MD. Viruses, Oncogenes and tumor suppressor genes. In: Tannock IF, Hill RP, editors. The basic science of oncology. New York: McGraw Hill; 1998. p. 79-105.</RefTotal>
</Reference>
<Reference refNo="50">
<RefAuthor>Tubbs RR</RefAuthor>
<RefAuthor>Hicks DG</RefAuthor>
<RefAuthor>Cook J</RefAuthor>
<RefAuthor>Downs-Kelly E</RefAuthor>
<RefAuthor>Pettay J</RefAuthor>
<RefAuthor>Hartke MB</RefAuthor>
<RefAuthor>Hood L</RefAuthor>
<RefAuthor>Neelon R</RefAuthor>
<RefAuthor>Myles J</RefAuthor>
<RefAuthor>Budd GT</RefAuthor>
<RefAuthor>Moore HC</RefAuthor>
<RefAuthor>Andresen S</RefAuthor>
<RefAuthor>Crowe JP</RefAuthor>
<RefTitle>Fluorescence in situ hybridization (FISH) as primary methodology for the assessment of HER2 Status in adenocarcinoma of the breast: a single institution experience</RefTitle>
<RefYear>2007</RefYear>
<RefJournal>Diagn Mol Pathol</RefJournal>
<RefPage>207-10</RefPage>
<RefTotal>Tubbs RR, Hicks DG, Cook J, Downs-Kelly E, Pettay J, Hartke MB, Hood L, Neelon R, Myles J, Budd GT, Moore HC, Andresen S, Crowe JP. Fluorescence in situ hybridization (FISH) as primary methodology for the assessment of HER2 Status in adenocarcinoma of the breast: a single institution experience. Diagn Mol Pathol. 2007;16(4):207-10. DOI: 10.1097/PDM.0b013e318064c72a</RefTotal>
<RefLink>http://dx.doi.org/10.1097/PDM.0b013e318064c72a</RefLink>
</Reference>
<Reference refNo="51">
<RefAuthor>Vanden Bempt I</RefAuthor>
<RefAuthor>Vanhentenrijk V</RefAuthor>
<RefAuthor>Drijkoningen M</RefAuthor>
<RefAuthor>Wlodarsaka I</RefAuthor>
<RefAuthor>Vandenerghe P</RefAuthor>
<RefAuthor>De Wolf-Peeters C</RefAuthor>
<RefTitle>Real-time reverse transcription-PCR and fluorescence in-situ hybridization are complementary to understand the mechanisms involved in HER-2/neu overexpression in breast carcinomas</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Histopathology</RefJournal>
<RefPage>431-41</RefPage>
<RefTotal>Vanden Bempt I, Vanhentenrijk V, Drijkoningen M, Wlodarsaka I, Vandenerghe P, De Wolf-Peeters C. Real-time reverse transcription-PCR and fluorescence in-situ hybridization are complementary to understand the mechanisms involved in HER-2/neu overexpression in breast carcinomas. Histopathology. 2005;46(4): 431-41. DOI: 10.1111/j.1365-2559.2005.02112.x</RefTotal>
<RefLink>http://dx.doi.org/10.1111/j.1365-2559.2005.02112.x</RefLink>
</Reference>
<Reference refNo="52">
<RefAuthor>Wilson CA</RefAuthor>
<RefAuthor>Cajulis EE</RefAuthor>
<RefAuthor>Green JL</RefAuthor>
<RefAuthor>Olsen TM</RefAuthor>
<RefAuthor>Chung YA</RefAuthor>
<RefAuthor>Damore MA</RefAuthor>
<RefAuthor>Dering J</RefAuthor>
<RefAuthor>Calzone FJ</RefAuthor>
<RefAuthor>Slamon DJ</RefAuthor>
<RefTitle>HER-2 overexpression differentially alters transforming growth factor-beta responses in luminal versus mesenchymal human breast cancer cells</RefTitle>
<RefYear>2005</RefYear>
<RefJournal>Breast Cancer Res</RefJournal>
<RefPage>R1058-79</RefPage>
<RefTotal>Wilson CA, Cajulis EE, Green JL, Olsen TM, Chung YA, Damore MA, Dering J, Calzone FJ, Slamon DJ. HER-2 overexpression differentially alters transforming growth factor-beta responses in luminal versus mesenchymal human breast cancer cells. Breast Cancer Res. 2005;7(6):R1058-79. DOI: 10.1186/bcr1343</RefTotal>
<RefLink>http://dx.doi.org/10.1186/bcr1343</RefLink>
</Reference>
<Reference refNo="53">
<RefAuthor>Witzel I</RefAuthor>
<RefAuthor>Thomssen C</RefAuthor>
<RefAuthor>Krenkel S</RefAuthor>
<RefAuthor>Wilczak W</RefAuthor>
<RefAuthor>Bubenheim M</RefAuthor>
<RefAuthor>Pantel K</RefAuthor>
<RefAuthor>Neumann R</RefAuthor>
<RefAuthor>Jänicke F</RefAuthor>
<RefAuthor>Müller V</RefAuthor>
<RefTitle>Clinical utility of determination of HER-2/neu and EGFR fragments in serum of patients with metastatic breast cancer</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>Int J Biol Markers</RefJournal>
<RefPage>131-40</RefPage>
<RefTotal>Witzel I, Thomssen C, Krenkel S, Wilczak W, Bubenheim M, Pantel K, Neumann R, Jänicke F, Müller V. Clinical utility of determination of HER-2/neu and EGFR fragments in serum of patients with metastatic breast cancer. Int J Biol Markers. 2006;21(3):131-40.</RefTotal>
</Reference>
<Reference refNo="54">
<RefAuthor>Wyllie AH</RefAuthor>
<RefTitle>The genetic regulation of apoptosis</RefTitle>
<RefYear>1995</RefYear>
<RefJournal>Curr Opin Genet Dev</RefJournal>
<RefPage>97-104</RefPage>
<RefTotal>Wyllie AH. The genetic regulation of apoptosis. Curr Opin Genet Dev. 1995;5(1): 97-104. DOI: 10.1016/S0959-437X(95)90060-8</RefTotal>
<RefLink>http://dx.doi.org/10.1016/S0959-437X(95)90060-8</RefLink>
</Reference>
<Reference refNo="55">
<RefAuthor>Yamamoto D</RefAuthor>
<RefAuthor>Senzaki H</RefAuthor>
<RefAuthor>Nakagawa H</RefAuthor>
<RefAuthor>Okugawa H</RefAuthor>
<RefAuthor>Gondo H</RefAuthor>
<RefAuthor>Tanaka K</RefAuthor>
<RefTitle>Detection of chromosomal aneusomy by fluorescence in situ hybridization for patients with nipple discharge</RefTitle>
<RefYear>2003</RefYear>
<RefJournal>Cancer</RefJournal>
<RefPage>690-4</RefPage>
<RefTotal>Yamamoto D, Senzaki H, Nakagawa H, Okugawa H, Gondo H, Tanaka K. Detection of chromosomal aneusomy by fluorescence in situ hybridization for patients with nipple discharge. Cancer. 2003;97(3):690-4. DOI: 10.1002/cncr.11091</RefTotal>
<RefLink>http://dx.doi.org/10.1002/cncr.11091</RefLink>
</Reference>
<Reference refNo="56">
<RefAuthor>Yang J</RefAuthor>
<RefAuthor>Zhou L</RefAuthor>
<RefAuthor>Hu R</RefAuthor>
<RefAuthor>Zhao J</RefAuthor>
<RefAuthor>Yao X</RefAuthor>
<RefAuthor>Long Q</RefAuthor>
<RefAuthor>Fan W</RefAuthor>
<RefAuthor>Chen L</RefAuthor>
<RefAuthor>Xing T</RefAuthor>
<RefTitle>[Expression of P62c-myc protein in breast cancer and its clinical significance]</RefTitle>
<RefYear>1998</RefYear>
<RefJournal>Hua Xi Yi Ke Da Xue Xue Bao</RefJournal>
<RefPage>399-401</RefPage>
<RefTotal>Yang J, Zhou L, Hu R, Zhao J, Yao X, Long Q, Fan W, Chen L, Xing T. [Expression of P62c-myc protein in breast cancer and its clinical significance]. Hua Xi Yi Ke Da Xue Xue Bao. 1998;29(4):399-401.</RefTotal>
</Reference>
<Reference refNo="57">
<RefAuthor>Yassin D</RefAuthor>
<RefAuthor>Kamel A</RefAuthor>
<RefAuthor>Taha H</RefAuthor>
<RefAuthor>Lotayef M</RefAuthor>
<RefAuthor>Hassan Y</RefAuthor>
<RefAuthor>Fawzy A</RefAuthor>
<RefAuthor>Akl S</RefAuthor>
<RefTitle>HER family expression in Egyptian breast cancer patients</RefTitle>
<RefYear>2003</RefYear>
<RefJournal>J Egypt Natl Canc Inst</RefJournal>
<RefPage>373-80</RefPage>
<RefTotal>Yassin D, Kamel A, Taha H, Lotayef M, Hassan Y, Fawzy A, Akl S. HER family expression in Egyptian breast cancer patients. J Egypt Natl Canc Inst. 2003;15(4):373-80.</RefTotal>
</Reference>
<Reference refNo="2">
<RefAuthor>Altman DG</RefAuthor>
<RefTitle></RefTitle>
<RefYear>1991</RefYear>
<RefBookTitle>Practical statistics for medical research</RefBookTitle>
<RefPage></RefPage>
<RefTotal>Altman DG. Practical statistics for medical research. London: Chapman and Hall; 1991.</RefTotal>
</Reference>
</References>
<Media>
<Tables>
<Table format="png">
<MediaNo>3</MediaNo>
<MediaID>3</MediaID>
<Caption>
<Pgraph>
<Mark1>Table 3: The level of HER-2/neu, c-myc and their relation to clinicopathological factors</Mark1>
</Pgraph>
</Caption>
</Table>
<Table format="png">
<MediaNo>4</MediaNo>
<MediaID>4</MediaID>
<Caption>
<Pgraph>
<Mark1>Table 4: Correlation between p53, c-myc and HER-2/neu in the malignant group</Mark1>
</Pgraph>
</Caption>
</Table>
<Table format="png">
<MediaNo>2</MediaNo>
<MediaID>2</MediaID>
<Caption>
<Pgraph>
<Mark1>Table 2: Positivity rate of HER-2/neu, c-myc and the % positivity of p53 and their relation to clinicopathological factors</Mark1>
</Pgraph>
</Caption>
</Table>
<Table format="png">
<MediaNo>1</MediaNo>
<MediaID>1</MediaID>
<Caption>
<Pgraph>
<Mark1>Table 1: The level of HER-2/neu, c-myc and P53 in benign and malignant groups</Mark1>
</Pgraph>
</Caption>
</Table>
<NoOfTables>4</NoOfTables>
</Tables>
<Figures>
<Figure format="png" height="498" width="562">
<MediaNo>1</MediaNo>
<MediaID>1</MediaID>
<Caption>
<Pgraph>
<Mark1>Figure 1: ROC curve of both HER-2/neu and c-myc. The area under the curve is 0.789 and 0.752 for HER-2/neu and c-myc, respectively.</Mark1>
</Pgraph>
</Caption>
</Figure>
<NoOfPictures>1</NoOfPictures>
</Figures>
<InlineFigures>
<NoOfPictures>0</NoOfPictures>
</InlineFigures>
<Attachments>
<NoOfAttachments>0</NoOfAttachments>
</Attachments>
</Media>
</OrigData>
</GmsArticle>
]]></plaintext></textgroup></pgraph></textblock></origdata></gmsarticle>