The relative prognostic significance of total cathepsin D and HER-2/neu oncogene amplification in breast cancer

Total tumor cathepsin D (TCD) levels were determined prospectively by a radioimmunometric assay in tumor cytosol of 858 primary breast cancer patients diagnosed between 1989–1991. In 581 of these patients, tumor HER-2/neu oncogene amplification was simultaneously determined. In a “training-set” of 313 patients, “high” TCD was associated with significantly shorter disease-free survival (DFS). For the whole group, there was no correlation between TCD and pathologic stage, number of axillary nodes with tumor deposits, tumor size, histologic type and grade, or hormone receptor levels. In the node-positive group, high TCD level was associated with HER-2/ neu amplification. After a median follow-up duration of 31 months, univariate analysis indicated that high TCD level was significantly associated with shorter DFS only in node-positive patients. The shorter DFS in association with high TCD levels was observed in both estrogen-receptor-positive and -negative patients. Cox multivariate analysis of DFS confirmed that high TCD level was predictive of shorter DFS in node-positive patients only. Because of the short duration of follow-up, the significance of TCD in overall survival was not determined. We conclude that high tumor TCD in node-positive patients is predictive of shorter DFS, and is often associated with HER-2/ neu amplification. The possibility exists that high tumor TCD may act in combination with HER-2/neu amplification to promote dissemination of metastases. © 1994 Wiley-Liss, Inc.     

The significance of intramammary nodes in primary breast cancer

The use of sentinel node biopsy in primary breast cancer raises many new controversies with regard to extra-axillary nodes. Three cases with intramammary nodes are discussed in relation to sentinel node biopsy.     

乳腺癌耐药蛋白在乳腺癌组织中的表达及与预后关系

目的研究乳腺癌耐药蛋白（Breast cancer resistance protein,BCRP）在乳腺癌组织中的表达,评估其在乳腺癌预后中的作用。方法采用免疫组织化学方法（Immunohistochemistry,IHC）检测47例手术切除的乳腺癌组织中BCRP的表达,并分析其与临床、病理特征的关系及对预后的影响。结果（1）BCRP在乳腺癌组织中的阳性表达率为55．3％（26／47例）,其中高表达者13例（27．7％）;（2）激素受体阳性者BCRP表达水平明显高于激素受体阴性者（P〈0．05）,BCRP表达与月经状况、肿瘤大小、腋淋巴结转移和组织分级均无关（P〉0．05）;（3）Kaplan—Meier生存分析结果表明BCRP和无病生存期相关（P〈0．05）,但和总生存期无关（P〉0．05）;（4）Cox单因素分析显示肿瘤大小和BCRP表达与无病生存期明显相关（P〈0．05）,而肿瘤大小和总生存期也明显相关（P〈0．05）;在多因素分析中BCRP表达仅和无病生存期明显相关,此外腋淋巴结转移与无病生存期和总生存期均明显相关（P〈0．05）。结论BCRP在乳腺癌组织中具有较高的表达水平,但与乳腺癌患者预后无关。     

Chromosomal rearrangements and oncogene amplification precede aneuploidization in the genetic evolution of breast cancer

Breast carcinoma is thought to arise because of multiple successive changes in the genome of the normal epithelial cells. However, little is known of the order of appearance of different types of genetic aberrations We studied the ERBB2 (Her-2/neu) and CCND1 (cyclin D1) oncogene amplification in flow cytometrically sorted diploid and nondiploid tumor cell populations by fluorescence in situ hybridization (FISH). The purity of the cell sorting was confirmed by static DNA image cytometry. Spectral karyotyping was used to define differences in a genome-wide manner between two distinctly different aneuploid cell clones found in each of two breast cancer cell lines. FISH indicated the presence of gene amplification both in diploid and nondiploid cell clones in 17 of the 21 amplification-containing tumors analyzed. The oncogene copy numbers remained unchanged throughout aneuploidization in 11 of 17 tumors. The remaining six tumors showed an increase in oncogene copy number as well as the number of chromosome 11 or 17 centromeres (the original location of CCNDI and ERBB2, respectively). Breast carcinoma cell lines MDA-157 and MDA-436 showed a significant number of chromosomal rearrangements in the near-diploid clones, which were present in duplicate in the corresponding aneuploid (polyploid) clones. These results indicate that ploidy shift, ie., aneuploidization, in breast cancer is a late genetic event which is preceded by both oncogene amplifications as well as many chromosomal rearrangements.     

FISH detection of HER-2/neu oncogene amplification in early onset breast cancer

HER-2/neu (c-erbB-2) gene amplification based on Southern blotting or immunohistochemistry has been shown to be predictive of poor outcome in breast cancer occurring in women over 40, but there is little data on the role of HER-2/neu in young women with breast cancer, many of whom may have inherited BRCA1 or other predisposing genes. The present study used fluorescent in situ hybridization (FISH) on archival specimens of breast cancer from 37 women under the age of 40 to evaluate the role of HER-2/neu amplification in this cohort, and to also evaluate the efficacy of FISH for quantifying amplification. The frequency of primary tumors with a greater than fourfold increase in gene copy number was found to be 38%, which is similar to the frequency of amplification reported in Southern blot studies in older women. However, the greater sensitivity of FISH enabled detection of low level amplification (more than 2 but less than 8 gene copies), which was found in an additional 30% of the tumors. Patients with low level amplification demonstrated a 54% recurrence rate, compared to 86% in those with high amplification and 17% in those with no amplification. HER-2/neu amplification appeared to be more prognostic of recurrence than nodal status, with 45% of node negative tumors recurring compared to 62% of those which were node positive, nor was tumor size predictive of recurrence in this cohort since tumors of 2 cm or less recurred in 44% of cases compared to 57% of those larger than 2 cm. Thus, this study demonstrates that FISH is a reproducible and sensitive technique for detecting HER-2/neu amplification, and that amplification of the oncogene is the strongest independent indicator of recurrence of breast cancer in young women.     

Relationship between oncogene amplification, aneuploidy and altered expression of p53 in breast cancer

Many studies have noted an association between amplification of single oncogenes and poor prognosis in breast cancer. We are investigating whether measurement of amplification in a larger number of proto-oncogenes increases the reliability of the prognostic information provided. As the first stage of this investigation, amplification (of c-erbB-2, cycD1, int-2, c-myc and MDM2), aneuploidy and altered expression of p53, which all indicate genetic instability, were studied in 117 primary breast adenocarcinomas. Amplification was correlated with aneuploidy (p=0.002) but not with altered expression of p53 even though the tumours with p53 overexpression were all aneuploid. Our results suggest that measurement of amplification is a potentially valuable prognostic factor.     

C-myc oncogene amplification in ductal carcinoma in situ of the breast

The c-myc oncogene is frequently activated in invasive breast cancer and has been associated with high nuclear grade, lymph node metastasis and poorer disease outcome. We have examined c-myc oncogene amplification using fluorescence in situ hybridization in a series of 96 pure DCIS. Additionally we assessed amplification and expression of the Her2 and bcl-2 oncogenes. The findings were compared with clinicopathological data, Ki-67 proliferative index and hormone receptor status. We observed c-myc oncogene amplification in 19 tumours (20%). These cases were significantly associated with an average of 38% higher proliferative activity (p = 0.045), a 43% larger tumour size (p = 0.029) and the otherwise rare micropapillary subtype (p = 0.0005). Concluding the c-myc oncogene appears to be involved in the development of a more aggressive phenotype of DCIS.     

Gamma-heregulin has no biological significance in primary breast cancer

British Journal of Cancer (2002) 86, 1362–1363. DOI: 10.1038/sj/bjc/6600245 www.bjcancer.com© 2002 Cancer Research UK     

Evaluation of the clinical significance of HER2 amplification by chromogenic in situ hybridisation in patients with primary breast cancer

BACKGROUND: The purpose of this study was to assess the clinical relevance of HER2 amplification by a novel chromogenic in situ hybridisation (CISH) technique in patients with primary breast cancer and to determine its relationship with other prognostic markers. MATERIALS AND METHODS: One hundred and seventy-three breast cancer patients with a mean follow-up duration of 75 months were reanalysed in this retrospective study. Expression of HER2 in tumour tissue samples was assessed by immunohistochemistry (IHC) and CISH. Discrepant cases and tumours presenting a HER2 2+ and 3+ staining with IHC were additionally analysed by fluorescence in situ hybridisation (FISH) to exclude false-positive results. RESULTS: HER2 overexpression and amplification was found in 24.3% and 19.1%, respectively. The clinico-pathological correlations revealed a significant association between positive HER2 status and standard prognostic factors including high tumour grade, large tumour size and absence of steroid hormone receptors. Univariate analysis indicated that HER2 overexpression and amplification were predictive for poor overall (OS) and disease-free survival (DFS). The same effect was also seen in the patient groups with node-negative as well as node-positive breast cancer. By multivariate analysis, HER2 alteration proved to be an indicator of poor prognosis, independent of tumour size, tumour grade, hormone receptor expression, nodal involvement and adjuvant therapy. CONCLUSION. HER2 expression, as assessed by CISH, is an independent marker for unfavourable prognosis in primary breast cancers.     

Follow-up study of HER-2/neu amplification in primary breast cancer

Amplification of the HER-2/neu oncogene was determined in 362 tumors from patients with primary breast cancer (185 node-positive patients and 177 node-negative patients). The overall amplification rate was 33% (30% for node-negative patients; 31% for patients with 1-3 positive nodes; 40% for patients with greater than 3 positive nodes). Gene copy number was not associated with axillary lymph node status, steroid receptor status, or patient age but was weakly correlated with the size of the primary tumor. Amplification of the HER-2/neu gene did not correlate with either disease-free or overall survival in univariate or multivariate analyses. The results were unambiguously negative for patients with node-negative disease. Although the univariate results for node-positive patients were marginally significant (P = 0.07), the significance was not retained in multivariate analyses. Thus, while HER-2/neu amplification may be biologically important in primary breast cancer, it will only be of marginal utility as a prognostic factor for predicting clinical outcome.     

Prognostic significance of serum IgE levels in primary breast cancer

Summary Serum IgE was measured in presurgical sera from 166 nonallergic women admitted to a comprehensive, multidisciplinary study of primary, operable breast cancer. During the follow-up period, which averaged 48 months, there were 71 recurrences. Patients were divided into two groups: those with IgE levels greater than the geometric mean value of 24 I.U. and those with levels less than the mean. The rate of tumor recurrence was significantly greater for the IgE > 24 group (p<0.03). IgE remained a significant prognostic indicator when evaluated by Cox regression analysis in conjunction with other known prognostic factors including: number of positive lymph nodes, clinical stage, menopausal status, estrogen receptor status, mitotic grade, tumor diameter, breast feeding history, and age of patient (p<0.015). IgE was not correlated with any of these known prognostic factors in individual analyses. We conclude that serum IgE level is a significant, independent prognostic indicator in primary breast cancer. Address for reprints: Dr H. Ownby, Michigan Cancer Foundation, 110 East Warren Avenue, Detroit, MI 48201, USA.     

Prognostic significance of estrogen receptor determination in primary breast cancer

The authors report on 767 consecutive primary Stage I-II breast cancer cases followed-up from 3 to 8 years. The estrogen receptor (ER) content was determined in all cases and did not influence the treatment choice. A correlation was attempted between ER and menstrual or pathological nodal status (N) or the 5-year disease-free survival (DFS). ER was correlated with menopausal status ER+ cases being more frequent in postmenopausal patients, whereas no correlation was observed between ER and nodal status. In absence of nodal involvement (N-) the prognosis was not influenced by the ER status. A significantly better DFS was evident for ER+ respect to ER- patients in the N+ series but such a correlation is questionable as the adjuvant treatment (hormone or chemotherapy) given to such patients may have influenced the DFS according to the ER status. According to the present study, ER determination should not be used as a discriminant in the performance of adjuvant postoperative treatment based on a prognostic judgment.     

The long term prognostic significance of c-erbB-2 in primary breast cancer.

The expression of the c-erbB-2 oncogene has been evaluated using an immunohistochemical technique with the 21N polyclonal antibody in paraffin embedded tissue from 465 patients treated between the years 1975-1981 for Stage I and II breast cancer. One hundred and four (22%) patients exhibited positive staining. This was not associated with any other variables. Expression of the oncogene was associated with significantly poorer survival which was independent of other tumour variables.     

Varying degrees of amplification of the N-ras oncogene in the human breast cancer cell line MCF-7

The oncogene N-ras has been found to be amplified (congruent to 20 copies) in the human breast carcinoma cell line MCF-7. The amplified sequences have been localized to a marker chromosome by in situ hybridization. Sublines of MCF-7, serially passaged in different laboratories, have marked variation in the degree of N-ras amplification. The differing degrees of amplification of N-ras are further evidence of heterogeneity within MCF-7 subclones. The phenomenon may not have general relevance for breast cancer, since other breast cancer cell lines and DNA from patient biopsies failed to show evidence of N-ras amplification.     

乳腺癌耐药蛋白在乳腺癌组织中的表达及其与预后的关系

目的:研究乳腺癌耐药蛋白(BCRP)在乳腺癌组织中的表达,评估其在乳腺癌预后中的作用。方法:采用免疫组织化学方法(IHC)检测60例手术切除的乳腺癌组织中BCRP的表达,并分析其与临床病理特征的关系及对预后的影响。结果:①BCRP在乳腺癌组织中的阳性表达率为35%(21/60例);②腋淋巴结或激素受体阳性者BCRP表达水平显著高于腋淋巴结阴性者和激素受体阴性者(P<0.05),BCRP表达与年龄、月经状况、肿瘤大小和组织学分级均无关(P>0.05);③Kaplan-Meier生存分析结果表明BCRP表达与无病生存期显著相关(P<0.05),但和总生存期无关(P>0.05);④Cox单因素和多因素分析都显示肿瘤大小、淋巴结转移和雌激素受体(ER)与无病生存期和总生存期显著相关(P<0.05),另外孕激素受体与总生存期(P<0.05)显著相关。结论:BCRP在乳腺癌组织中具有一定的表达水平,与乳腺癌患者的无病生存期有关,而与总生存期无关。     

Risk of breast cancer according to the status of HER-2/neu oncogene amplification.

We examined risk factors for breast cancer after subdividing cases based on the presence of HER-2/neu oncogene amplification in their tumors. Data were from the Carolina Breast Cancer Study, a population-based, case-control study of 577 invasive breast cancer patients, diagnosed during 1993-1996 and ages 20-74 years, and 790 controls frequency-matched on race and age. Information on breast cancer risk factors was obtained from structured personal interviews. About 20% of paraffin-embedded tissues from the breast cancers of cases were identified as positive for HER-2/neu amplification (HER-2/neu+) by differential PCR. Early age at menarche, higher waist:hip ratio, and family history of breast or ovarian cancer were associated with elevated odds ratios (ORs) for both HER-2/neu+ and HER-2/neu- breast cancers. Breastfeeding for at least 1 year was inversely associated with HER-2/neu+ breast cancer [OR, 0.3; 95% confidence interval (CI), 0.1-0.7] more so than HER-2/neu- breast cancer (OR, 0.8; 95% CI, 0.5-1.2). Most of the remaining risk factors had ORs around 1.0 for both HER-2/neu+ and HER-2/neu- breast cancers, although a few exhibited possible associations with one disease subtype in analyses stratified by menopausal status. These study results suggest that most recognized breast cancer risk factors do not operate through HER-2/neu amplification in breast carcinogenesis. Differential effects of long-term breastfeeding by HER-2/neu amplification status have been observed in earlier studies and are provocative; however, the direction and magnitude of the associations have not been consistent.     

肺癌C-myc基因扩增的临床意义

目的与方法:应用聚合酶链反应(PCR)技术对56例肺癌石蜡包埋标本进行C-myc基因的研究.结果:C-myc基因扩增与组织学类型、临床病理分期及淋巴结转移有密切关系(P<0.05),C-myc基因扩增者,分化程度较低,临床病理分期较晚,淋巴结转移多.同时还发现C-myc基因扩增与预后有明显关系(P<0.05),C-myc基因扩增者预后较差.结论:提示C-myc基因可以作为判断肺癌预后的检测指标之一.     

乳腺癌组织中survivin、c-erbB-2基因的表达与预后的关系

目的:探讨乳腺癌组织中survivin基因和c-erbB-2基因的表达与预后的关系。方法:用免疫组化S-P法检测80例乳腺癌组织中survivin、c-erbB-2的表达,分析其与腋淋巴结转移和5年无病生存率之间的关系。结果:survivin基因在乳腺癌中阳性表达率为68.75%(55/80)、c-erbB-2基因阳性表达率为36.25%(29/80),均与腋淋巴结转移正相关,与5年生存率负相关(P<0.05);survivin和c-erbB-2表达与肿瘤病理类型、发病年龄、临床分期无明显相关性(P>0.05);survivin和c-erbB-2基因表达相互呈正相关(P<0.05)。结论:凋亡抑制基因survivin和原癌基因c-erbB-2可能在乳腺癌的发生、发展过程中起重要作用,联合检测能更好地判断乳腺癌的预后。     

In vivo amplification and rearrangement of c-myc oncogene in human breast tumors

We have studied the genomic organization of cellular myc (c-myc) proto-oncogene in 48 human primary breast tumors. Two types of alterations (amplification and rearrangement) were observed in 27 (56%) of the tumors studied. The c-myc proto-oncogene appeared to be amplified 2- to 15-fold in the DNA of 20 tumors (41%). Non-germ line c-myc-related fragments (rearrangements) of variable size were detected in 7 primary breast tumors (6 malignant, 1 benign); 4 of these tumors presented both rearrangement and amplification, and the other 3 presented rearrangement only. The majority of the tumors analyzed were invasive ductal adenocarcinomas; 58% of these showed c-myc locus genetic alterations. Although the c-myc alterations described here do not appear to correlate with the aggressive behavior of primary breast tumors, they seem to be associated with development of breast carcinoma.