Clusterin和Syk在大肠癌中的表达及意义

[目的]分析凋亡相关因子Clusterin和脾酪氨酸激酶（Syk）与大肠癌生物学行为的关系。[方法]应用免疫组化法检测80例大肠癌组织和癌周正常组织中Clusterin和Syk表达水平。[结果]Clusterin在正常大肠组织中表达率为10.00%（8/80）,在大肠癌组织中的表达率为71.25%（57/80）,两者比较有显著性差异（P〈0.01）。Syk在正常大肠组织中表达率为92.50%（74/80）,在大肠癌组织中的表达率为35.00%（28/80）,两者比较有显著性差异（P〈0.01）。Clusterin表达与大肠癌分化程度、浸润深度、TNM分期、淋巴结转移有关（P〈0.05）,Syk表达水平与大肠癌浸润深度、TNM分期、淋巴结转移有关（P〈0.05）。[结论]大肠癌组织中Clusterin高表达,Syk低表达,两者可作为判断大肠癌恶性潜能的生物学指标。     

HIF-1α和VEGF在大肠癌中的表达及其相关性研究

[目的]探讨HIF-1α和VEGF在大肠癌浸润、转移过程中的表达及其相互作用机制。[方法]采用免疫组化SP法检测90例大肠癌组织及其相应癌旁组织中HIF-1α和VEGF的表达。[结果]癌旁组织中HIF-1α的表达均为阴性,大肠癌组织中的阳性表达率为73.33%（66/90）,显著高于癌旁组织,差异有统计学意义（P〈0.05）。HIF-1α在大肠癌中阳性表达率与临床分期、浸润深度和淋巴结转移有关（P〈0.05）,与大肠癌分化程度无关（P〉0.05）。癌旁组织中VEGF的表达均为阴性,在大肠癌组织中的阳性表达率为70.00%（63/90）,显著高于癌旁组织,差异有统计学意义（P〈0.05）。VEGF在大肠癌中阳性表达率与临床分期、浸润深度和淋巴结转移有关（P〈0.05）,与大肠癌分化程度无关（P〉0.05）。大肠癌组织中HIF-1α和VEGF蛋白表达具有正相关性（r=0.592,P〈0.05）。[结论]HIF-1α与VEGF的表达率均随着大肠癌浸润深度、淋巴结转移和临床分期的发展而增高,共同参与了大肠癌的发生、发展,可作为大肠癌浸润和转移的重要判定指标。HIF-1α可能通过上调VEGF的蛋白表达,促进大肠癌血管生成而促进大肠癌的转移,抑制HIF-1α的表达可能是治疗大肠癌的新手段。     

EphA2蛋白表达与结直肠癌浸润性及微血管生成的关系

目的：探讨酪氨酸蛋白激酶受体EphA2表达与大肠癌浸润及其与大肠癌微血管生成之间的关系。方法：应用免疫组化方法检测70例大肠癌标本和相应正常大肠黏膜组织中EphA2蛋白的表达情况,分析E-phA2表达与大肠癌临床病理学因素及微血管生成之间的关系。结果：EphA2蛋白在癌组织中的表达明显高于相应正常大肠黏膜组织（P〈0.001）。EphA2蛋白高表达与癌分化程度、生长方式、浸润深度和肿瘤体积相关（P〈0.05）,而与患者的年龄、性别、淋巴结和血行性转移无关（P〉0.05）。免疫组化结果还发现,肿瘤组织内微血管内皮细胞也有EphA2受体蛋白表达。CD34染色后大肠癌的微血管密度（microvessel density,MVD）与EphA2表达水平有显著相关,EphA2阳性表达强度高的肿瘤区域有较高的微血管密度。结论：E-phA2蛋白的高表达与大肠癌的发生和浸润有关。     

环氧化酶-2在乳腺癌中高表达的临床意义

[目的]探讨环氧化酶-2（COX-2）在乳腺癌组织中的表达及其与临床病理因素和预后的相关性。[方法]应用免疫组织化学方法检测60例乳腺癌石蜡包埋组织中COX-2、HER-2／neu和血管内皮生长因子（VEGF）的表达。[结果]60例乳腺癌组织中COX-2的高表达率为383％（23／60）。在淋巴结转移阳性组和阴性组以及不同临床分期病人COX-2的表达差异有显著性（P〈0．05）；COX-2的表达与HER-2／neu、VEGF表达密切相关（P〈0．01）。COX-2和HER-2／neu共同表达的患者TNM分期较晚．COX-2低表达的患者总生存期较长，与高表达患者的总生存期有显著性差异（P〈0．05）。[结论]乳腺癌组织中COX-2的高表达可提示乳腺癌恶性程度高和预后不良．COX-2和HER-2／neu同时高表达预后更差．     

MCM2在大肠癌组织中的表达及其临床意义

目的：研究微型染色体维持蛋白（minichromosome maintenance proteins,MCMs）家族中的一成员MCM2蛋白在大肠癌中的表达情况及其临床意义。方法：用免疫组化SP法检测50例大肠癌组织、20例大肠腺瘤组织、10例正常肠粘膜中MCM2的表达,分析MCM2与大肠癌临床病理因素之间关系。结果：10例正常大肠粘膜中未见MCM2表达,50例大肠癌组织中MCM2阳性表达率为54%（27/50）,两者差异有统计学意义（P〈0.05）。MCM2表达与大肠癌分化程度、淋巴结转移有统计学意义（P〈0.05）。与患者年龄、性别、肿瘤大小、浸润深度、临床Ducks分期无关（P〉0.05）。结论：MCM2表达是反映大肠癌生物学行为的有价值指标。     

大肠癌组织Lumican基因及蛋白的表达及临床意义

目的：检测人大肠癌组织中Lumican基因及蛋白的表达,探讨其与大肠癌患者临床病理指标及预后之间的关系。方法：收集外科手术切除并经病理证实的27例新鲜大肠癌组织标本,以同一患者远离肿瘤的正常大肠组织为对照,采用实时荧光定量PCR法检测Lumican基因表达,剩余组织采用石蜡包埋用于免疫组化检测Lumican蛋白。另从我院病理科收集同期37例大肠癌组织蜡块,采用免疫组化SP法共检测64例大肠癌组织、27例正常大肠黏膜组织中Lumican蛋白表达。结果：27例新鲜大肠癌组织及对应正常大肠组织中Lumican的表达有显著差异性（ P〈0.001）,Lumican mRNA在大肠癌组织表达明显增高[以正常组织为1,癌组织Lumican基因表达上调（5.35±0.66）倍]。64例大肠癌组织Lumican蛋白阳性表达率为73.44%（47/64）,显著高于远离肿瘤的正常组织表达率0%（0/27）（P〈0.05）。Lumican表达与肿瘤的病理分级、Dukes分期、淋巴结是否转移有关（P〈0.001）,而与患者性别、年龄、病变部位无关（P〉0.05）。27例大肠癌标本Lu-mican蛋白量与mRNA相对表达量呈正相关（r=0.123）,但无统计学意义（P〉0.05）。结论：人大肠癌组织Lumican mRNA及蛋白均表达升高,Lumican高表达与大肠癌的临床病理特征及侵袭、转移特性密切相关,在大肠癌发生、发展过程可能起着重要作用。Lumican可作为判断大肠癌生物学行为的重要指标之一。     

DLL4和VEGF蛋白在大肠癌中的表达及其临床意义

目的：检测大肠癌组织中DLL4和VEGF的表达,探讨其与临床病理参数的关系、临床意义及两者的相关性。方法：采用免疫组化Envision法检测60例结直肠癌及21例癌旁正常组织中DLL4、VEGF的表达情况。结果：DLL4和VEGF在大肠癌组织中表达的阳性率分别为75.0%、61.67%,明显高于癌旁正常组织（P〈0.05）。DLL4及VEGF蛋白在大肠癌中的表达与肿瘤浸润深度、淋巴结转移、远处转移及TNM分期密切相关（P〈0.05）;二者在大肠癌组织中的表达与患者性别、年龄、肿瘤部位、肿瘤大小及组织学分化程度无关（P〉0.05）。DLL4阳性表达病例中的VEGF表达率明显高于DLL4阴性表达的病例,DLL4和VEGF蛋白表达水平呈明显正相关（r=0.257,P〈0.05）。结论：DLL4和VEGF在大肠癌和正常大肠组织中的表达差异有统计学意义,DLL4在大肠癌组织中的表达上调与VEGF有关,它们可能共同调控肿瘤新生血管发生,参与大肠癌的侵袭和转移,从而影响患者的预后。     

大肠癌组织c-met与VEGF的表达及意义

[目的]研究肝细胞生长因子（HGF）受体c-met与血管内皮生长因子（VEGF）在大肠癌组织中的表达,探讨两者与肿瘤微血管生成及临床病理特征间的关系。[方法]采用免疫组化法检测56例（其中41例癌旁正常黏膜）大肠癌组织中c-met、VEGF表达以及MVD参数。[结果]c-met、VEGF在大肠癌组织中的阳性表达率分别为67.8%和53.6%;在癌旁正常黏膜组织中的阳性率分别为12.2%和14.6%（P〈0.001）。c-met、VEGF的阳性标本MVD值与其阴性标本之间有显著性差异（P〈0.05）。VEGF表达与肿瘤浸润深度、Dukes’分期及淋巴结转移情况有关（P〈0.05）,而与分化程度、年龄、性别无关（P〉0.05）。c-met表达与肿瘤分化程度、Dukes’分期、浸润深度及淋巴结转移情况有关（P〈0.05）,而与性别、年龄无关（P〉0.05）。c-met表达与VEGF表达趋于一致。[结论]c-met、VEGF在大肠癌组织中的表达显著高于癌旁黏膜组织。c-met与VEGF相互作用共同促进肿瘤血管生成。     

Livin蛋白在人大肠癌组织中的表达及临床意义

目的检测凋亡抑制因子Livin在大肠肿瘤组织中的表达,探讨Livin蛋白与大肠癌不同临床病理特征及预后的关系。方法采用免疫组织化学MaxVision^TM二步法检测Livin蛋白在178例大肠癌组织、60例癌旁组织及30例正常大肠组织中的表达情况。结果 178例大肠癌组织中Livin蛋白阳性率为65.7%（117/178）,高于癌旁组织（8.33%）及正常肠组织（0%）（P〈0.001）;Livin蛋白表达与肿瘤浸润深度、Duke’s分期、临床分期以及远处转移关系密切（P〈0.05）。经Cox回归模型多因素分析,示Livin蛋白表达是大肠癌患者长期生存的独立危险因素。结论人大肠癌组织中存在Livin蛋白表达,Livin蛋白表达与大肠癌患者的预后有关。     

大肠癌组织中Cox-2和FasL的表达及意义

目的:探讨Cox2和FasL蛋白在大肠癌组织中的表达、相互关系及临床意义。方法应用免疫组化SP法检测60例大肠癌、20例大肠腺瘤及20例正常大肠组织Cox2和FasL蛋白的表达情况。结果:1)大肠癌组织、大肠腺瘤和正常大肠组织Cox2表达阳性率分别为78.3%(47/60)、55.0%(11/20)和25.0%(5/20),Cox2在大肠癌组织中阳性表达率显著高于腺瘤组织及正常大肠组织阳性表达率,P=0.000。大肠癌、大肠腺瘤和正常大肠组织阳性表达率分别为86.7%(52/60)、50.0%(10/20)和20.0%(4/20)FasL在大肠癌组织中阳性表达率显著高于腺瘤组织及正常组织,P=0.000。2)Cox2和FasL的表达与肿瘤组织学分级及肿瘤生长部位无关Cox2与肿瘤Duke’s分期和淋巴结转移有相关性,P值分别为0.0.25和0.006。Cox2的表达与肿瘤大小有关,P=0.005。3)大肠癌组织中Cox2与FasL蛋白的表达为正相关性,r=0.627,P=0.000。结论:大肠癌组织中Cox2和FasL高表达,两者之间的表达强度有等级相关性。Cox2蛋白可通过诱导FasL蛋白的表达上调,增加大肠癌细胞侵袭力,从而成为其促进癌细胞浸润、转移的途径之一。     

Quantitative association between HER-2/neu and steroid hormone receptors in hormone receptor-positive primary breast cancer

BACKGROUND: HER-2/neu, which encodes a receptor tyrosine kinase, is amplified and overexpressed in 20%-25% of human breast cancers. Such tumors are often resistant to hormone therapy. Despite a general inverse association between HER-2/neu amplification/overexpression and estrogen receptor (ER) and/or progesterone receptor (PR) expression, a fraction of patients are both HER-2/neu- and hormone receptor (HR)-positive. The efficacy of hormone therapy in this group is currently a matter of debate. To better understand the relationship between HER-2/neu positivity and HR expression, we analyzed HER-2/neu, ER, and PR as continuous variables in breast cancer cell lines and two cohorts of primary breast cancer patients. METHODS: HER-2/neu and ER/PR expression was analyzed by enzyme-linked immunosorbent assay (ELISA) and enzyme immunoassay (EIA), respectively, in 14 human breast cancer cell lines, some of which had been transfected with the HER-2/neu gene. For the clinical study population, HER-2/neu protein levels were assessed by ELISA (cohort A, n = 665), and HER-2/neu gene copy number was determined using fluorescence in situ hybridization (cohort B, n = 894). ER/PR expression was analyzed by EIA (cohort A) or radioligand binding (cohort B). Associations between HER-2/neu and ER/PR expression were analyzed using Spearman's rho correlation and the chi-square test, and absolute levels were compared using the Mann-Whitney U test. All statistical tests were two-sided. RESULTS: HR-positive human breast cancer cell lines transfected with the HER-2/neu gene expressed statistically significantly lower levels of ER and PR than parental lines. In the clinical cohorts, levels of HER-2/neu overexpression and gene amplification were inversely correlated with ER/PR levels (Cohort A [n = 112]: for ER, r = -0.34, P<.001; for PR, r = -0.24, P =.010. Cohort B [n = 188]: for ER, r = -0.39, P<.001; for PR, r = -0.26, P<.001). Among patients with HR-positive tumors, HER-2/neu-positive tumors had statistically significantly lower ER/PR levels than HER-2/neu-negative ones (Cohort A: for ER, median = 25 fmol/mg [interquartile range [IQR] = 13-78] versus median = 38.5 fmol/mg [IQR = 17-99] and P =.031; for PR, median = 35 fmol/mg [IQR = 12-119] versus median = 88.5 fmol/mg [IQR = 22-236] and P<.001. Cohort B: for ER, median = 44 fmol/mg [IQR = 13-156] versus median = 92 fmol/mg [IQR = 35-235] and P<.001; for PR, median = 36 fmol/mg [IQR = 13-108] versus median = 84 fmol/mg [IQR = 24-250] and P<.001). Patients with higher levels of HER-2/neu overexpression or amplification had statistically significantly lower levels of ER/PR than patients with lower levels of HER-2/neu overexpression or amplification. CONCLUSION: Because absolute HR levels are strongly related to response to hormone therapy in primary and advanced breast cancer, reduced ER/PR expression may be one mechanism to explain the relative resistance of HER-2/neu-positive:HR-positive tumors to hormone therapy.     

Overexpression of HER-2/neu is associated with poor survival in advanced epithelial ovarian cancer

Previous studies have suggested that overexpression of HER-2/neu oncogene occurs in 15-40% of breast cancers and that overexpression is associated with poor prognosis. In the present report, we have used an immunohistochemical technique involving a monoclonal antibody specifically reactive with the external domain of HER-2/neu to study expression of HER-2/neu in frozen sections of normal ovary and advanced epithelial ovarian cancer. The intensity of staining for HER-2neu was always moderate or less (0-2+) in normal ovarian epithelium. Among 73 ovarian cancers, 50 (68%) had staining similar to that for normal ovarian epithelium (0-2+) while 23 (32%) stained heavily (3+). Survival of the 23 patients with high HER-2/neu expression (median, 15.7 months) was significantly worse (P = 0.001) than that of the 50 patients (median, 32.8 months) with normal HER-2/neu expression. In addition, patients whose tumors had high HER-2/neu expression were significantly less likely to have a complete response to primary therapy (P less than 0.05) or have a negative second-look laparotomy when serum CA 125 levels were normal preoperatively (P less than 0.05). These findings suggest that HER-2/neu deserves further evaluation as a prognostic marker in epithelial ovarian cancer.     

Correlation of p27 protein expression with HER-2/neu expression in breast cancer

Strong expression of human epidermal growth factor receptor 2 (HER-2)/neu in breast cancer has been associated with poor prognosis. Reduced expression of p27(Kip1), a cyclin-dependent kinase inhibitor, correlates with poor clinical outcome in breast cancer. In this study, we provide a correlation between these two important prognostic markers in patients with breast cancer. Breast tumor screening using immunohistochemistry indicated that downregulation of p27 correlated with HER-2/neu overexpression in studying 11 normal breast tissues and 51 primary breast carcinomas. We found HER-2/neu protein overexpression in 20 (41%) of 49 breast cancers and low p27 protein expression in 47 (92%) of 51 breast cancers. All 20 (100%) of the tumors that overexpressed HER-2/neu had low levels of p27 protein product; this correlation was statistically significant (P = 0.035). Decreasing p27 expression correlated with increasing HER-2/neu activity. Our results suggest that one function of the HER-2/neu product is to downregulate p27 expression in breast cancer. This study may be significant in selecting patients for HER-2/neu antibody therapy in the future. Mol. Carcinog. 30:169--175, 2001.     

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.     

HER-2/neu and p53 expression versus tamoxifen resistance in estrogen receptor-positive, node-positive breast cancer.

PURPOSE: An association between the overexpression of proto-oncogene HER-2/neu and resistance to tamoxifen in estrogen receptor (ER)-positive primary and metastatic breast cancer has been suggested. We examine a possible interaction between HER-2/neu or p53 expression and tamoxifen effectiveness in patients with ER-positive, node-positive disease treated with cyclophosphamide, doxorubicin, and fluorouracil in a large adjuvant chemotherapy trial (Cancer and Leukemia Group B [CALGB] 8541). Tamoxifen assignment was not randomized-physician discretion was used for premenopausal and postmenopausal women. Trial protocol then specified assignment to postmenopausal women with ER-positive tumors, although not all took tamoxifen. PATIENTS AND METHODS: CALGB 8541 assessed HER-2/neu expression in patients with ER-positive disease by immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) and amplification by differential polymerase chain reaction (PCR). IHC assessed expression of p53. Univariate and multivariate proportional hazards models assessed tamoxifen-HER-2/neu status interactions and tamoxifen-p53 status interactions. RESULTS: HER-2/neu status was available for 651 patients with ER-positive disease; 650, 608, and 353 patients were assessed by IHC, PCR, and FISH, respectively. Approximately one half received tamoxifen. Reduction in risk of disease recurrence or death resulting from tamoxifen was approximately 37% (32% with overexpression and 39% with normal expression of HER-2/neu; n = 155 by IHC). The tamoxifen-HER-2/neu status interaction was not significant in multivariate analysis of all three HER-2/neu assessment methods. Tamoxifen-p53 interaction did not significantly predict outcome. CONCLUSION: Disease-free and overall survival benefit of tamoxifen in patients with ER-positive, node-positive breast cancer does not depend on HER-2/neu or p53 status. Our data suggest that neither HER-2/neu nor p53 expression should be used to determine assignment of tamoxifen.     

HER-2 receptor expression, localization, and activation in colorectal cancer cell lines and human tumors

The HER-2/neu oncogene encodes a 185 kD protein that is phosphorylated upon ligand binding to other HER/erbB members and regulates cell growth and differentiation. Given that HER-2 receptor blockade can inhibit the growth of colon cancer cell lines and tumor xenografts, we investigated the frequency, localization and phosphorylation status of HER-2 in colon cancer cell lines and in human tumors. Protein expression was analyzed in relation to mRNA levels, HER-2 amplification, and clinicopathological variables. Colon cancer cell lines constitutively expressed HER-2 proteins and none showed HER-2 amplification by fluorescence in situ hybridization. Cell fractionation and immunoblotting showed HER-2 in both the membrane and cytosolic compartments. Primary colorectal carcinomas (n = 96) and their metastases (n = 25) were examined by immunohistochemistry. Strong membrane HER-2 staining was detected in 5 (5%) of primaries and in 3 (12%) metastases (p = 0.36). Membrane but not cytoplasmic localization was strongly associated with HER-2 gene amplification (p = 0.007). Cytoplasmic HER-2 staining was found in 61 (63.5%) of primary tumors and localization was confirmed by immunoelectron microscopy that also showed plasma membrane HER-2. Using real-time quantitative RT-PCR, HER-2 mRNA was increased in tumors with membrane compared to cytoplasmic staining (r = 0.66, p = 0.001). Cytoplasmic HER-2 was associated with tumor differentiation (p = 0.018), but not other clinicopathological variables. By immunoblotting, heterogeneity was seen in HER-2 levels with downregulation in 4 of 7 tumors relative to normal epithelia that uniformly expressed HER-2. Phosphorylated HER-2 was detected in approximately 50% of tumors and in normal mucosa. In conclusion, HER-2 is expressed constitutively in colon cancer cell lines and demonstrates relatively distinct localization patterns in human tumors. Strong membrane immunoreactivity is associated with high levels of HER-2 mRNA and gene amplification whereas cytoplasmic HER-2 is detected frequently and seems to be a marker of tumor differentiation.     

COX-2及Her2/neu在结直肠癌旁组织中表达意义

目的 检测结直肠癌和旁癌组织中的COX-2及Her2/neu的表达情况,探究COX-2及Her2/neu表达与结直肠癌临床各指标的关系,并探讨其临床意义.方法 选取结直肠癌患者200例,收集各研究对象的结直肠癌病理标本和癌旁正常组织各一份,应用免疫组织化学法(S-P法),检测结直肠癌组织和癌旁正常组织中COX-2及Her2/neu蛋白质表达.结果 结直肠癌中Her2/neu表达情况:蛋白质表达阳性占86.0%,明显高于对照组Her/neu阳性表达(24.0%)(P<0.05).结直肠癌中Her2/neu表达与Dukes分期和肝转移相关(P<0.05).结直肠癌中COX-2表达阳性率为89.0%,而癌旁正常组织中其阳性表达率为32.0%,两者差异具有统计学意义(P<0.05),过表达与肿瘤肝转移和Dukes分期有关(P<0.05),与Her2/neu表达有正相关性(P<0.05).结论 采用免疫组织化学法探究蛋白质的表达,结肠癌COX-2及Her2/neu蛋白质的表达率高于癌旁正常组织;COX-2及Her2/neu蛋白质的表达与肝转移和Dukes有关,而且它们之间的表达具有正相关性(P<0.05).