Expression of the HER-2/neu proto-oncogene in serous ovarian neoplasms.

BACKGROUND. It is unclear whether HER-2/neu proto-oncogene expression in ovarian epithelial neoplasms is related to prognosis. METHODS. The authors performed immunohistochemical stains on 20 serous tumors of low malignant potential (STLMP) in Stages I and II and 19 serious carcinomas in the same stages. They used three different commercial antibodies to make comparisons. RESULTS. Two of four Stage I STLMP in patients who experienced disease progression showed positive staining for the gene product, whereas none of seven Stage I nonprogressive STLMP showed positive staining. Five of the six Stage III nonprogressive STLMP showed positive staining, whereas none of three Stage III STLMP that progressed showed positive staining. Three carcinomas (one Stage I and two Stage III) also showed positive staining. CONCLUSIONS. Expression of HER-2/neu may be associated with high stage in serous ovarian neoplasms, but it is not likely to identify the small fraction of patients with STLMP who will experience disease progression.     

Expression of c-erb B-2/neu proto-oncogene in human prostatic cancer tissues and cell lines.

Both amplification and overexpression of c-erb B-2/neu have been associated with the progression and possible prognosis of a number of human cancers. In this study, we demonstrated that c-erb B-2/neu may also play an important role in human prostate cancer. Our conclusion is based on the following observations: (1) A monoclonal antibody raised against a peptide sequence from the C-terminal domain of the human c-erb B-2/neu gene product reacted positively with 68.7% (11 of 16) of the human prostatic cancer tissue extracts analyzed by western blot procedure. These results were supported by the immunohistochemical staining of the prostatic cancer specimens; 80% (12 of 15) showed positive staining, primarily around the plasma membranes of the prostatic cancer cells. c-erb B-2/neu oncoprotein was not detectable in normal prostate tissues (five examined by immunohistochemical staining and three by western blotting) or in human benign prostatic hyperplasia (two examined by immunohistochemical staining and six by western blotting) and was expressed less abundantly with lower intensity in "normal" human prostate tissues adjacent to cancerous prostate tissue (5 of 12 examined by immunohistochemical staining). We observed no evidence of c-erb B-2/neu gene amplification in 10 fresh human prostatic cancer specimens examined by Southern blotting and in the cultured human prostatic cancer cell lines PC-3, DU-145, and LNCaP. (2) The c-erb B-2/neu protein was detected in both androgen-receptor-positive (LNCaP) and -negative (PC-3 and DU-145) human prostate cancer cell lines. Positive immunostaining of c-erb B-2/neu protein was found to be associated predominantly with the plasma membranes of PC-3 cells, but was also found to be widespread in the cytoplasmic region of the LNCaP cells and in the perinuclear region of the DU-145 cells. (3) Like prostate-specific antigen (PSA) expression, c-erb B-2/neu mRNA expression was also positively regulated by androgen in androgen-receptor-positive LNCaP cells in vitro and LNCaP tumors in vivo. When LNCaP tumors were grown in castrated male hosts, levels of c-erb B-2/neu and PSA mRNA expression decreased initially, but rebounded at 3 wk to levels comparable to those expressed by tumors maintained in intact adult male hosts.     

胃腺癌组织HER-2/neu基因表达及其临床意义

目的:探讨胃腺癌HER-2/neu基因表达与临床病理参数和术后5年生存率的关系。方法:显色原位杂交(CISH)检测55例胃腺癌组织HER-2/neu基因表达。结果:55例胃腺癌组织HER-2/neu基因扩增5例(阳性率9.09%);阳性病例组织学类型均为肠型胃癌(Lauren分型);HER-2/neu基因扩增患者生存率明显低于HER-2/neu基因不扩增患者(P=0.019)。结论:HER-2/neu基因是导致胃腺癌术后5年生存率降低的重要因子,显色原位杂交法检测胃腺癌中HER-2/neu基因扩增状态,可指导trastuzumab靶向治疗胃腺癌。     

Expression of ras proto-oncogene proteins in normal human tissues

The expression of ras proto-oncogenes in normal human tissues was studied by immunohistochemical staining and by immunoblotting using monoclonal antibodies. We detected p21ras protein in almost every fetal and adult tissue, but the level varied significantly among cell types. In some cell lineages, immature cells capable of proliferation contain more p21ras than do mature cells. By contrast, certain fully differentiated cells, such as neurons and the epithelial cells of endocrine glands, express abundant p21ras. Among mammalian tissues the highest level of ras protein was detected in brain. Crude synaptosomal membrane preparations from rat brain contain substantially more p21ras than do plasma membranes from rat liver. The observed distribution of p21ras suggests a role for these proteins both in cellular proliferation and in certain specialized cellular functions.     

Expression of HER-2/neu in testicular tumors

BACKGROUND: Although the overexpression of the Epidermal Growth Factor Receptor 2 (EGFR-2, HER-2/neu, c-erbB-2) in malignancies might predict chemoresistance and poor prognosis, its clinical relevance has not been widely studied and determined in testicular tumors. PATIENTS AND METHODS: Since teratomas are relatively chemoresistant tumors, we evaluated the HER-2/neu receptor status of 28 primary testicular tumors (7 pure teratomas, 21 mixed germ cell tumors containing teratomatous components) using a standardized immunohistochemical method (HercepTest Kit). RESULTS: Seven (25%) out of 28 non-seminomatous germ cell tumors showed HER-2/neu positivity. The teratomatous components of mixed GCTs showed HER-2/neu overexpression in 5 cases. Three of the 5 choriocarcinoma components of mixed tumors overexpressed HER-2/neu. In one case (teratoma + choriocarcinoma) both components showed HER-2/neu overexpression. No HER-2/neu overexpression was detected in other, less differentiated histological subtypes. Among the HER-2/neu-positive cases, 3 patients are in complete remission, 3 patients are in partial remission and one patient died after primary chemotherapy. CONCLUSION: Twenty-five percent of the non-seminomatous germ cell tumors which contain teratomatous components overexpress HER-2/neu protein. The overexpression is restricted to the more differentiated histotypes. Further molecular investigations and clinicopathological studies are necessary to determine the correlation between HER-2/neu overexpression and clinical resistance of testicular tumors.     

Expression of the c-erbB-3 protein in normal human adult and fetal tissues.

c-erbB-3 is a member of the type I family of growth factor receptors which includes the epidermal growth factor (EGF) receptor and c-erbB-2. Whereas for EGF receptor and c-erbB-2 the expression patterns in normal tissues are well documented, there is currently little information available about the sites of c-erbB-3 expression. In order to examine the normal tissue distribution of c-erbB-3, polyclonal antibodies were raised against eight synthetic peptides corresponding to distinct sites on the intracellular domain of c-erbB-3. Of these, three produced antibodies which reacted with a 160-kDa protein on immunoblots of human embryonal cells (293 cells) transfected with the cDNA encoding c-erbB-3, and two of the three antibodies immunoprecipitated a protein of similar size from the same cells. These antibodies were used for immunochemical staining of a wide variety of normal human adult and fetal tissues employing formalin-fixed, paraffin-embedded material. The c-erbB-3 protein was identified in cells of the gastrointestinal, reproductive, respiratory and urinary tracts as well as the skin, endocrine and nervous system in a distribution distinctly different from that observed for EGF receptor and c-erbB-2. The level of expression of the mRNA for c-erbB-3 was also examined in extracts of a selection of fetal tissues. In general the sites of mRNA and protein expression were concordant.     

甲状腺癌HER—2／neu癌蛋白和mRNA表达的研究

「目的」观察人类甲状腺癌中ＨＥＲ－２／ｎｅｕ癌蛋白和ｍＲＮＡ的表达及其关系。「方法」应用免疫组化和同位素点杂交技术对新鲜冰冻甲状腺组织ＨＥＲ－２／ｎｅｕ癌蛋白和ｍＲＮＡ的表达进行研究。「结果」２１例甲状腺乳头状癌中１４例ＨＥＲ－２ｅｎｕ癌蛋白表达阳性,而其他类型肿瘤和非肿瘤组织均未见ＨＥＲ－２／ｎｅｕ ｍＲＮＡ表达研究显示,乳头状癌和淋巴结转移灶中,ｍＲＮＡ的表达水平明显高于其他肿瘤和非肿瘤组织。本研究还证明石蜡包埋的乳头状癌标本中ＨＥＲ－２／ｎｅｕ癌蛋白表达率明显低于新鲜标本。「结论」ＨＥＲ－２／ｎｅｕ蛋白的表达是甲状腺乳头状癌的一个特征性改变,ＨＥＲ－２ｎｅｕ蛋白的表达与ｍＲＮＡ的表达相一致,ＨＥＲ－２ｎｅｕ蛋白检测的稳定性取决于组织标本的质量。     

Expression of HER-2/neu in Oral Squamous Cell Carcinoma

Background: HER-2/neu is a member of the human epidermal growth factor (HER) family of transmembrane tyrosine kinases, which is significantly associated with the pathogenesis of various cancer types. The aim was to evaluate the expression of HER-2/neu in oral squamous cell carcinoma (OSCC) as a potential biomarker to target antigens for specific immunotherapy in OSCC. Methods: One hundred and forty histologically diagnosed OSCC cases were identified. Four to five-micrometer thick formalin-fixed, paraffin-embedded tumor sections were stained with Haematoxylin and Eosin (H and E). Histological grade was assessed according to WHO/Broders classification, while tumors were staged according to the American Joint Committee on Cancer (AJCC) TNM classification from stage I to IV. Immunohistochemistry was performed by using Rabbit monoclonal antibody against HER-2/neu (EP700Y, cell marquee and diluted 1:50). FISH was performed on positive cases using Vysis PathVysion HER-2 DNA probe (Abbott USA). Probes consist of LSI HER gene spectrum orange and control probe CEP 17 spectrum green. Results: In this study, males were mostly effected (64.3%) with buccal mucosa (49%) to be the commonly involved site for OSCC. Majority of cases were moderately differentiated (62.1%) and 50.7% tumors were Stage IV. HER-2/neu was found to be positive (2+) in one case of OSCC, however weak to moderate complete membrane staining was observed in >10% of the tumor cells. One hundred and thirty nine cases were HER-2/neu negative. FISH analysis of HER-2/neu positive cases also showed gene amplification (Her2-neu/ CEp 17 = 225/33 = 7.2). Conclusions: The study showed disparity in the expression of HER-2/neu in OSCC, which is due to multiple reasons. Therefore therapy against HER-2/neu in OSCC is debatable.     

HER-2/neu is a tumor rejection target in tolerized HER-2/neu transgenic mice

HER-2/neu (neu-N) transgenic mice, which express the nontransforming rat proto-oncogene, develop spontaneous focal mammary adenocarcinomas beginning at 5-6 months of age. The development and histology of these tumors bears a striking resemblance to what is seen in patients with breast cancer. We have characterized the immunological responses to HER-2/neu (neu) in this animal model. neu-positive tumor lines, which were derived from spontaneous tumors that formed in neu-N animals, are highly immunogenic in parental, FVB/N mice. In contrast, a 100-fold lower tumor challenge is sufficient for growth in 100% of transgenic animals. Despite significant tolerance to the transgene, neu-specific immune responses similar to those observed in breast cancer patients can be demonstrated in neu-N mice prior to vaccination. Both cellular and humoral neu-specific responses in transgenic mice can be boosted with neu-specific vaccination, although to a significantly lesser degree than what is observed in FVB/N mice, indicating that the T cells involved are less responsive than in the nontoleragenic parental strain. Using irradiated whole-cell and recombinant vaccinia virus vaccinations we are able to protect neu-N mice from a neu-expressing tumor challenge. T-cell depletion experiments demonstrated that the observed protection is T cell dependent. The vaccine-dependent neu-specific immune response is also sufficient to delay the onset of spontaneous tumor formation in these mice. These data suggest that, despite tolerance to neu in this transgenic model, it is possible to immunize neu-specific T cells to achieve neu-specific tumor rejection in vivo. These transgenic mice provide a spontaneous tumor model for identifying vaccine approaches potent enough to overcome mechanisms of immune tolerance that are likely to exist in patients with cancer.     

HER-2/neu in bladder carcinoma

A total of 66 bladder cancer patients were studied to verify possible relationships between HER-2/neu alterations and pathological characteristics, and to define a poor prognosis patient subgroup with respect to time to recurrence, time to progression and survival. Tumor and healthy tissue specimens were analyzed for HER-2/neu DNA amplification and protein overexpression by Southern and Western blot techniques and evaluated statistically. 13% of cases were amplified and 39% were overexpressed. HER-2/neu alterations were not significantly associated with pathological staging or tumor grading. Multifocal tumors had a higher percentage and overexpression with respect to monofocal tumors. Actuarial analyses did not show a significant statistical correlation between HER-2/neu amplification and overexpression and clinical outcome. Clinical evaluation of HER-2/neu status showed that this gene is not related to tumor relapse, progression or patient survival.     

HER-2/neu基因在鼻咽癌中的表达及其临床意义

目的：探讨HER－2／neu基因在鼻咽癌中的表达及其临床意义。方法：采用免疫组化法测定HER－2／neu基因蛋白的表达。结果：69例鼻咽癌病人中52％（36／69）有HER－2／neu基因蛋白表达。这种过表达与肿瘤分期、局部淋巴结和远处脏器转移以及5年总生存期、3年无病生存期均无相关性。同一病例原发病灶和复发或转移灶HER－2/neu基因蛋白的表达较一致。HER－2／neu基因蛋白过表达对鼻咽癌化疗的反应性不产生影响。结论：在鼻咽癌,HER－2／neu基因蛋白过表达的临床意义不前,HER-2/neu基因蛋白能否成为治疗新的靶点还有待进一步研究。     

HER-2/neu, c-Myc and cyclin-A in human breast cancer

To better understand the role of HER-2/neu, c-myc and cyclin-A which seem to be activated in different steps of tumor cell growth, we analyzed a series of breast cancers from patients subjected to radical mastectomy with regard to HER-2/neu amplification (N=171) and expression of HER-2/neu (N=114), c-myc (N=31) and cyclin-A (N=71). Molecular evaluation demonstrated that HER-2/neu was amplified in 20% of cases and overexpressed in 27%, and its alterations were associated with a higher proliferative activity (H-3-Tdr-labeling index), although not statistically significant in patients without lymph node metastases, c-myc was overexpressed in 16% of cases and was weakly correlated to proliferation activity. Cyclin-A was overexpressed in 15% of cases and was significantly correlated to the percentage of the H-3-Tdr labeled cell fraction (p=0.002). Cyclin-A alterations were also significantly associated with well-differentiated tumors, suggesting that this gene could be involved in the control of the normal cell cycle rather than to cell proliferation during tumor growth.     

HER-2 therapy. HER-2/neu diagnostics in breast cancer

HER-2/neu status of the primary breast cancer (PBC) is determined by immunohistochemistry and fluorescent in situ hybridization. Because of a variety of technical factors, however, the PBC may not accurately reflect the metastatic tumor in terms of HER-2/neu status. Recently published guidelines recommend that tumors be defined as HER-2/neu positive if 30% or more of the cells are 3+. Circulating levels of the HER-2 extracellular domain can be measured in serum using a test cleared by the US Food and Drug Administration, and increased serum HER-2/neu levels to above 15 ng/ml can reflect tumor progression. Studies comparing tissue HER-2/neu status of the PBC and HER-2/neu levels above 15 ng/ml in metastatic breast cancer patients are also reviewed.     

Pre-existent immunity to the HER-2/neu oncogenic protein in patients with HER-2/neu overexpressing breast and ovarian cancer

Immunomodulatory strategies, such as antibody therapy and cancer vaccines, are increasingly being considered as potential adjuvant therapies in patients with advanced stage breast cancer to either treat minimal residual disease or prevent relapse. However, little is known concerning the incidence and magnitude of the pre-existent breast cancer specific immune response in this patient population. Using the HER-2/neu oncogenic protein as a model, a well-defined tumor antigen in breast cancer, we questioned whether patients with advanced stage HER-2/neu overexpressing breast and ovarian cancers (III/IV) had evidence of pre-existent immunity to HER-2/neu. Forty-five patients with stage III or IV HER-2/neu overexpressing breast or ovarian cancer were evaluated for HER-2/neu specific T cell and antibody immunity. Patients enrolled had not received immunosuppressive chemotherapy for at least 30 days (median 5 months, range 1–75 months). All patients were documented to be immune competent prior to entry by DTH testing using a skin test anergy battery. Five of 45 patients (11%) were found to have a significant HER-2/neu specific T cell response as defined by a stimulation index 2.0 (range 2.0–7.9). None of eight patients who were HLA-A2 had a detectable IFN secreting T-cell precursor frequency to a well-defined HER-2/neu HLA-A2 T cell epitope, p369-377. Three of 45 patients (7%) had detectable HER-2/neu specific IgG antibodies, range 1.2–8.9g/ml. These findings suggest that patients with advanced stage HER-2/neu overexpressing breast and ovarian cancer can mount a T cell and/or antibody immune response to their tumor. However, in the case of the HER-2/neu antigen, the pre-existent tumor specific immune response is found only in a minority of patients.     

HER-2/neu diagnostics in breast cancer

HER-2/neu status of the primary breast cancer (PBC) is determined by immunohistochemistry and fluorescent in situ hybridization. Because of a variety of technical factors, however, the PBC may not accurately reflect the metastatic tumor in terms of HER-2/neu status. Recently published guidelines recommend that tumors be defined as HER-2/neu positive if 30% or more of the cells are 3+. Circulating levels of the HER-2 extracellular domain can be measured in serum using a test cleared by the US Food and Drug Administration, and increased serum HER-2/neu levels to above 15 ng/ml can reflect tumor progression. Studies comparing tissue HER-2/neu status of the PBC and HER-2/neu levels above 15 ng/ml in metastatic breast cancer patients are also reviewed.     

Amplification and expression of the c-erb B-2/neu proto-oncogene in human bladder cancer

The c-erb B-2/neu gene encodes a cell-surface glycoprotein with extensive homology to, but distinct from, the epidermal growth-factor receptor. In this study, we compared the c-erb B-2/neu gene amplification and expression of tissue specimens obtained from the bladders of normal controls and patients with high-grade transitional cell bladder carcinoma. Southern blot analysis of DNAs from 24 patients and 5 controls showed 2 cases of c-erb B-2/neu gene amplification in patients and none in controls. Western blot analysis demonstrated that c-erb B-2/neu was expressed in 67.6% (23/34) of patient specimens but in none of the controls (0/5). This finding agreed with the result of immunohistochemical staining, which showed that tissue from 74.3% (26/35) of the patients and none of the controls (0/7) showed positive immunofluorescence staining. This is the first report suggesting that c-erb B-2/neu gene amplification may be associated with human bladder carcinogenesis.     

The expression of HER-2/neu gene in colon cancer tissues and its clinical significance

Objective

The article aims to detect the expression of HER-2/neu gene in colon cancer tissues and adjacent tissues, to analyze the relationship between different pathologic types and clinical features, also to invest the distribution of patients with positive expression of HER-2 gene.

Methods

The expression of HER-2 gene in the 223 samples with colon cancer was detected by immunochemical approach. The expression of HER-2 gene in colon cancer tissues and adjacent tissues and different pathologic types was analyzed by χ² test. The correlation between the expression of HER-2 gene and clinical features was analyzed by Spearman.

Results

The number of positive expression of HER-2 gene in colon cancer tissues and adjacent tissues were 74 and 0 respectively, the difference has statistical significance. The number of papillary or tubular adenocarcinoma was 182, among them, 60 cases were positive expression. The number of mucinous adenocarcinoma was 41, among them, 14 cases were positive expression. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of differentiation and depth of invasion, which has no statistical significance. However, the expression of HER-2/neu gene has correlation with metastasis of lymph node and Dukes stage, which has statistical significance. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The correlated coefficient index was 0.320 and 0.320 respectively. In the 74 patients with positive expression of HER-2 gene, 59.4% of them were 60–74 years old. And there was 97.3% of the patients without family history of adenocarcinoma.

Conclusion

The expression of HER-2/neu gene in colon cancer tissues was higher than in adjacent tissues. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of differentiation and depth of invasion, but has correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene with age of 60–74 years old and without family history of adenocarcinoma was higher than other groups.     

Generation of T-cell immunity to the HER-2/neu protein after active immunization with HER-2/neu peptide-based vaccines.

PURPOSE: The HER-2/neu protein is a nonmutated tumor antigen that is overexpressed in a variety of human malignancies, including breast and ovarian cancer. Many tumor antigens, such as MAGE and gp100, are self-proteins; therefore, effective vaccine strategies must circumvent tolerance. We hypothesized that immunizing patients with subdominant peptide epitopes derived from HER-2/neu, using an adjuvant known to recruit professional antigen-presenting cells, granulocyte-macrophage colony-stimulating factor, would result in the generation of T-cell immunity specific for the HER-2/neu protein. PATIENTS AND METHODS: Sixty-four patients with HER-2/neu-overexpressing breast, ovarian, or non-small-cell lung cancers were enrolled. Vaccines were composed of peptides derived from potential T-helper epitopes of the HER-2/neu protein admixed with granulocyte-macrophage colony-stimulating factor and administered intradermally. Peripheral-blood mononuclear cells were evaluated at baseline, before vaccination, and after vaccination for antigen-specific T-cell immunity. Immunologic response data are presented on the 38 subjects who completed six vaccinations. Toxicity data are presented on all 64 patients enrolled. RESULTS: Ninety-two percent of patients developed T-cell immunity to HER-2/neu peptides (stimulation index, 2.1 to 59) and 68% to a HER-2/neu protein domain (stimulation index range, 2 to 31). Epitope spreading was observed in 84% of patients and significantly correlated with the generation of a HER-2/neu protein-specific T-cell immunity (P =.03). At 1-year follow-up, immunity to the HER-2/neu protein persisted in 38% of patients. CONCLUSION: The majority of patients with HER-2/neu-overexpressing cancers can develop immunity to both HER-2/neu peptides and protein. In addition, the generation of protein-specific immunity, after peptide immunization, was associated with epitope spreading, reflecting the initiation of an endogenous immune response. Finally, immunity can persist after active immunizations have ended.