HER-2/neu蛋白表达与胃癌临床病理特征及预后关系的研究

探讨胃癌组织中人表皮生长因子受体(HER)-2/neu蛋白的表达与患者临床病理特征及预后的关系。选取本院肿瘤科收治的98例胃癌确诊患者肿瘤组织标本及40例患者癌旁组织行免疫组化染色,观察两种组织标本中HER-2/neu蛋白表达情况,并分析其与胃癌患者临床病理特征和预后的关系。胃癌组织中HER-2/neu蛋白阳性表达率显著高于癌旁组织(38.78%比0,P0.05)。胃癌组织中HER-2/neu蛋白阳性表达主要与肿瘤淋巴结转移、远处转移、浸润深度、TNM分期、Lauren分型有关(P0.05)。HER-2/neu蛋白阴性表达患者的3年生存率显著高于阳性表达患者(58.33%比28.95%,χ~2=8.066,P0.05)。胃癌组织HER-2/neu蛋白阳性表达患者的中位生存时间显著低于阴性表达患者(17.9个月比28.6个月,χ~2=10.565,P0.05)。胃癌组织中HER-2/neu蛋白的表达与患者临床病理特征及远期预后密切相关。     

HER-2/neu siRNA对人胶质瘤细胞株T98G侵袭性的抑制作用

目的 探讨人表皮生长因子受体-2(HER-2/neu)特异性siRNA对高表达HER-2/neu的人胶质瘤细胞株T98G侵袭性的影响及其机制.方法 用脂质体介导终质量浓度50 nmol/LHER-2/neu siRNA转染人胶质瘤细胞株T98G,实时定量聚合酶链反应(real-time PCR)和免疫印迹法检测转染后HER-2/neu mRNA和蛋白表达变化.Transwell体外侵袭实验检测终质量浓度25、50、100 nmol/L HER-2/neu siRNA转染T98G后细胞侵袭能力的变化.免疫印迹法检测转染后细胞基质金属蛋白酶-9(MMP-9)蛋白表达变化.明胶酶谱法检测转染后MMP-9活性变化.结果 终质量浓度50 nmol/L HER-2/neu siRNA转染T98G细胞后HER-2/neu mRNA和蛋白表达为对照组的(29.3±6.2)%和(13.1±8.9)%(P＜0.01).终质量浓度25、50、100 nmol/L HER-2/neu siRNA转染后细胞侵袭抑制率分别为(37.7 ±7.2)%、(65.0±10.1)%和(69.7±9.3)%,三者与对照组的差异均有统计学意义(P＜0.01).与对照组比较,转染组细胞MMP-9蛋白表达减少(49.9±10.7)%,酶活性下降(55.6±12.7)%(P＜0.01).结论 HER-2/neu siRNA转染人胶质瘤细胞株T98G后明显抑制细胞的侵袭能力,其机制与MMP-9蛋白表达和活性显著受到抑制有关.     

TIP30/CC3基因在乳腺癌中表达的研究

目的观察TIP30/CC3基因在乳腺癌组织中的表达状况,分析其与乳腺癌相关临床病理指标间的关系。方法应用半定量RT-PCR检测40例乳腺癌组织与40例正常乳腺组织TIP30/CC3基因的表达情况。结果TIP30/CC3基因在乳腺癌组织及正常乳腺组织中均有表达,在40例乳腺癌组织中,TIP30/CC3基因呈低表达,35%（14/40）为阳性;而在正常组织中TIP30/CC3基因呈高表达,85%（34/40）为阳性,两者相比有显著性差异（P〈0.01）。TIP30/CC3基因的表达与C-erbB-2（χ^2=8.864,P=0.003）、P53（χ^2=10.989,P=0.001）呈显著相关;与乳腺癌患者绝经状态（χ^2=0.218,P=0.641）、ER（χ^2=1.283,P=0.257）、PR（P=1.000）、肿瘤大小（χ^2=0.584,P=0.445）以及肿瘤的淋巴结转移（χ^2=0.165,P=0.685）无关。结论TIP30/CC3基因在乳腺癌的发病中发挥重要作用,可能为乳腺癌综合治疗提供依据。     

血清HER-2/neu胞外域在复发转移性乳腺癌患者解救化疗中的临床意义

目的 探讨血清HER-2/neu胞外域(ECD)在复发转移性乳腺癌患者解救化疗中的临床意义.方法 酶联免疫吸附试验检测健康志愿者、良性乳腺疾病患者各27例的血清HER-2/neu ECD水平及27例复发转移性乳腺癌患者化疗前后的血清HER-2/neu ECD水平,结合其临床资料进行统计学分析.结果 健康志愿者与乳腺良性疾病患者血清HER-2/neu ECD水平均低于15μg/L,差异无统计学意义(P>0.05);复发转移性乳腺癌患者化疗前血清HER-2/neu ECD的平均浓度为(47.30±109.06)μg/L,显著高于健康志愿者及乳腺良性疾病患者(P<0.01).血清HER-2/neu ECD水平与原发肿瘤HER-2/neu的表达成正相关(r=0.752,P<0.01).解救化疗有效的患者,化疗3周后血清HER-2/neu ECD水平较化疗前明显降低(P<0.05),化疗3周后与化疗6周后血清HER-2/neu ECD水平差异无统计学意义(P>0.05).化疗前HER-2/neu ECD水平与患者总生存期呈负相关(r=-0.601,P<0.01),而与无进展生存期无明显相关(r=-0.374,P>0.05).化疗前血清HER-2/neu ECD>15 ug/L与HER-2/neu ECD<15 ug/L的患者比较,总生存率前者低于后者(P<0.05),无进展生存率两者间差异无统计学意义(P>0.05).结论 血清HER-2/neu ECD水平的监测,有助于早期发现乳腺癌的复发转移,指导解救化疗方案的选择,评估患者的疗效及预后.     

ER、PR、HER-2/neu与乳腺癌新辅助化疗有效率的关系研究

目的探讨乳腺癌患者ER、PR、HER-2/neu的表达及其与新辅助化疗有效率的关系及乳腺癌新辅助化疗敏感性的预测指标。方法采用免疫组化的方法检测160例乳腺癌新辅助化疗病人化疗前后ER、PR、HER-2/neu的表达情况,并评估其与乳腺癌新辅助化疗有效率的关系。结果160例乳腺癌新辅助化疗病例中,ER、PR阴性率、HER-2/neu过度表达率分别为35.63%,24.38%,33.13%,ER( )组与ER(-)组、PR( )组与PR(-)组及HER-2/neu过度表达组与非过度表达组化疗有效率差异均有统计学意义(χ2>3.875,P<0.05);新辅助化疗前后ER、PR的阳性率和HER-2/neu过度表达率的改变无统计学意义。结论ER或PR阴性的乳腺癌病人对化疗更敏感,化疗后获益更多,而HER-2/neu过度表达的病人化疗不敏感,ER、PR和HER-2/neu可作为乳腺癌新辅助化疗敏感性的预测指标;新辅助化疗不改变ER、PR及HER-2/neu的表达水平。     

HER-2/neu过表达与雄激素非依赖性前列腺癌Gleason分级和临床分期的相关性研究

目的:检测HER-2/neu基因在雄激素依赖与雄激素非依赖性前列腺癌(PCa)组织中的表达情况,推测这一基因在雄激素非依赖性PCa发生中的意义。方法:收集30例雄激素依赖性PCa标本(雄激素依赖组)和24例激素非依赖性PCa标本(雄激素非依赖组),采用免疫组化方法检测HER-2/neu基因的蛋白表达,对照临床分期、Gleason分级进行分析。结果:HER-2/neu在雄激素依赖性PCa标本中的阳性表达率为10%,在雄激素非依赖性PCa的表达为33%;Gleason分级≤7分病例中HER-2/neu阳性表达率显著低于Gleason分级>7分病例组(14.29%vs26.92%,P<0.01);临床分期>T2病例中HER-2/neu阳性表达率显著高于≤T2病例组(34.62%vs7.14%,P<0.01)。结论:HER-2/neu在雄激素非依赖性PCa标本中阳性表达率高,而且随临床分期和Gleason分级的升高其阳性表达率升高。     

TIP30/CC3基因在食管癌中的研究进展

目的:探讨TIP30/CC3基因在食管癌中的表达.方法:用免疫组化S-P法检测64例食管癌组织和72例正常食管组织中TIP30/CC3基因的表达.结果: 72例正常食管组织中有63例(87.5％) TIP30/CC3基因表达.64例食管鳞癌中有38例(59.38％) TIP30/CC3基因表达,差异具有显著性意义(P＜0.05);TIP30/CC3基因在高分化、中分化、低分化食管鳞癌中的表达率分别为78.13％ (25/32)、 42.11％ (8/19)、 38.46％(5/13),比较差异有统计学意义(P＜0.05);TIP30/CC3基因的表达在淋巴结转移组中为39.13％ (9/23),明显低于未发生淋巴结转移组70.73％ (29/41),(P＜0.05),差异有统计学意义.结论:食管癌TIP30/CC3基因的表达与肿瘤的侵袭性有关,可作为预后不良的指标.     

应用荧光原位杂交技术检测骨肉瘤患者人表皮生长因子2受体基因扩增状态

目的 应用荧光原位杂交技术检测骨肉瘤人表皮生长因子2受体(HER-2/neu)基因是否存在扩增.方法 以HER-2/neu及17号染色体为探针,应用荧光原位杂交技术分析23例骨肉瘤病例HER-2/neu基因扩增情况,以HER-2/neu基因扩增阳性的乳腺癌病例作为阳性对照.结果 FISH检测23例骨肉瘤标本,无一例出现HER-2/neu基因扩增.结论 骨肉瘤中出现HER-2/neu基因扩增的频率极低,骨肉瘤患者可能不适合应用以HER-2/neu基因为靶点的靶向性药物.     

siRNA干扰叉头框C2对乳腺癌中肿瘤干细胞标志物CD44的影响

目的：探讨siRNA干扰叉头框C2（FOXC2）对乳腺癌中肿瘤干细胞（CSC）标志物CD44 mRNA及蛋白表达的影响。 方法：常规培养乳腺癌MCF-7细胞株，再通过乳腺球形成实验培养、筛选乳腺癌CSC；将FOXC2-siRNA或阴性对照siRNA慢病毒载体转染至乳腺癌CSC，同时将转染载体的CSC作为空白对照。通过real time RT-PCR和Western blot检测FOXC2-siRNA对FOXC2的干扰效应，再通过real time RT-PCR和Western blot分别检测CD44 mRNA和蛋白在各组细胞中的表达，结果均以空白对照的表达量作为参照进行分析。 结果：从MCF-7细胞株中成功培养出乳腺癌CSC。与转染阴性对照siRNA的乳腺癌CSC比较，转染FOXC2-siRNA的乳腺癌CSC中FOXC2 mRNA和蛋白表达均明显降低（P=0.00509；P=0.00001），同时CD44 mRNA及蛋白的表达也均明显降低（P=0.00848；P=0.00218）。 结论：干扰乳腺癌CSC中FOXC2基因的表达能够抑制乳腺癌CSC中CD44 mRNA及蛋白的表达，FOXC2信号转导通路可能通过调控CD44来介导乳腺癌CSC的增殖、分化。     

免疫组化和荧光原位杂交检测HER-2与乳腺癌病理参数的相关性

目的:探讨免疫组化和荧光原位杂交检测HER-2与乳腺癌患者病理参数的相关性。方法:采用免疫组化二步法和荧光原位杂交法检测80例乳腺癌组织中HER-2表达情况,分析其与临床病理参数的关系。结果:乳腺癌组织中HER-2蛋白表达阳性率为56.25%(45/80),而基因扩增率为30%(24/80)。HER-2蛋白的表达和HER-2基因扩增均与乳腺癌组织学分级、淋巴结转移密切相关(均P0.05),而与患者年龄、肿瘤大小无关(P0.05)。结论:乳腺癌组织中HER-2蛋白表达和基因扩增与肿瘤分化程度密切相关,具有重要的临床意义。     

Prognostic value of HER-2/neu and p53 expression in node-positive breast cancer. HER-2/neu effect on adjuvant tamoxifen treatment

HER-2/neu and p53 expression, conventional clinical and pathologic prognostic factors, were evaluated in a retrospective series of 283 node-positive breast cancer patients. Overexpression was determined by immunohistochemistry in formalin-fixed paraffin-embedded tissue blocks. Twenty one percent were HER-2/neu positive and 40% p53 positive. HER-2/neu expression was related to axillary lymph node metastasis (P=0.014), inflammatory infiltrates (P=0.004), and the absence of oestrogen (ER) (P=0.0026) and progesterone (P=0.01) receptors (PR). p53 expression was related to lymph node involvement (P=0.03), necrosis (P=0.036), absence of ER (P=0.028) and PR (P=0.065). p53 was not associated with outcome. HER-2/neu was an unfavourable prognostic factor for disease-free (DFS) (P=0.05) and overall survival (OS) (P=0.02) in univariate analysis. Multivariate analysis showed that the number of involved axillary nodes (P<0.00001), age (P=0.004), grade (P=0.04), and PR (P=0.04) were independent predictors for OS. ER-positive patients treated with adjuvant tamoxifen had shorter DFS and OS when they were HER-2/neu positive.     

Concordance of the hormone receptors and correlation of HER-2/neu overexpression of the metachronous cancers of contralateral breasts

The primary objective of this study was to evaluate the relative prevalence of estrogen receptor-negative contralateral breast cancer to the first primary cancer and to assess the correlation between the relative overexpression of HER-2/neu in the first primary cancer and contralateral breast cancer. A total of 144 women diagnosed with cancers in contralateral breasts were identified from the Henry Ford Health System tumor registry. Data were retrieved from electronic databases and medical records. Women were dichotomized into users and nonusers of tamoxifen. Hormone receptors were scored as positive or negative. HER-2/neu overexpression, assessed by immunohistochemistry, was scored as 0, 1(+), 2(+), or 3(+). Concordance between hormone receptors of the two cancers was low (kappa = 0.27, p = 0.06). Stratification of women by tamoxifen therapy yielded an almost fivefold increase in the proportion of estrogen receptor-negative cancers among the users, while the proportion of cancers expressing no estrogen receptor remained the same among the nonusers (39.6% versus 40.6%). Matched, archived, paraffin-embedded specimens of the first and contralateral breast cancers were available for 57 women. The correlation between the relative overexpression of HER-2/neu between the first primary and the contralateral breast cancer was 0.4 (p = 0.002). The higher prevalence of estrogen receptor-negative contralateral breast cancer among tamoxifen users concurs with previous reports. The biological mechanism for this observation is not understood; however, it has been proposed that tamoxifen inhibits the proliferation of estrogen receptor-positive breast cancer cells, while estrogen receptor-negative cells may continue to grow because of selective pressure. The correlation between HER-2/neu overexpression in the matched first primary and contralateral breast cancers was statistically significant, suggesting that the diagnosis of HER-2/neu overexpression in contralateral breast cancer is associated with HER-2/neu overexpression in the first primary cancer.     

Histopathologic characteristics predicting HER-2/neu amplification in breast cancer

The HER-2/neu gene is a proto-oncogene that is amplified in 10-30% of breast cancers. New drugs for targeted therapy, such as Herceptin, are effective for patients with HER-2/neu-positive tumors, making it necessary to have a noncostly and accurate method to assess HER-2/neu status. We studied the correlation of findings made by fluorescent in situ hybridization (FISH) and immunohistochemistry (IHC) staining and the possibility of combining IHC and other clinicopathologic characteristics of breast tumors to predict FISH-determined HER-2/neu status. The clinicopathologic characteristics analyzed were the size of the tumor, p53, lymph-vascular invasion, estrogen/progesterone receptors (ER/PR), tumor grade, axillary lymph node status, and patient age. A total of 199 cases of invasive breast cancer studied at the UCLA Pathology Laboratory during 2003 were included in this study. Tumors with IHC 0, 1+, 2+, and 3+ scores were found to be FISH positive in 3.5%, 6.4%, 25.7%, and 81.5% of the respective groups. Our study showed a strong association between the FISH-negative and IHC scored 0 and 1+ tumors, suggesting that the FISH test may not be necessary in these cases (p<0.0001). Although the concordance between IHC 3+ and FISH positive is high, 18% of the patients with overexpression of HER-2/neu fail to show gene amplification by FISH. HER-2/neu positivity was found to be proportionally associated with increasing grade in infiltrating ductal carcinoma (p<0.0001). p53-positive tumors are more likely to be HER-2/neu amplified (p=0.0003). Tumors that are negative for ER/PR are also associated with HER-2/neu positivity by FISH (31.15%, p=0.0016). FISH-determined HER-2/neu status is not associated with histologic type, tumor size, nodal status, lymph-vascular invasion, or patient age.     

HER-2/neu Expression in Primary Breast Cancer With Sentinel Lymph Node Metastasis

Background Amplification of the protein product of the HER-2/neu oncogene in primary breast cancer specimens is associated with an adverse prognosis. We hypothesized that overexpression of HER-2/neu would predict metastases to the sentinel lymph nodes (SLNs). Methods A retrospective review of a prospective nonrandomized evaluation of 1055 clinically node-negative breast cancer patients undergoing 1063 SLN biopsies was performed. HER-2/neu analysis was performed by immunohistochemistry and, in selected cases, by fluorescence in situ hybridization. Clinical, demographic, surgical, radiological, and pathologic data were analyzed by using generalized estimating equations logistic regression models. Results Two hundred thirty-two (23.6%) of 985 operations in which the SLN was found at operation resulted in positive nodes. In a multivariate analysis, size (P < .0001) and HER-2/neu overexpression (P = .026) were independent predictors of SLN metastasis. Conclusions Size is a known predictor of SLN metastasis in the modern SLN era, as it was in the pre-SLN eras. HER-2/neu was found to be significantly predictive of SLN metastasis in our study. We anticipate a future when even the relatively minor procedure of SLN biopsy might be avoided with the predictive information gained from studying the pathology and molecular markers of primary breast cancers.     

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目的　研究HER 2 /neu(c erbB 2 )在结、直肠恶性肿瘤中的表达及其与肿瘤分期的相关性。方法　收集 2 0 0 2年 1月至 2 0 0 3年 5月治疗的 2 8例结、直肠恶性肿瘤的临床资料。按Dukes分期标准进行分期 ,其中A、B期共 17例 (17/2 8,6 0 7% ) ,C、D期 11例 (11/2 8,39 3% )。采用HER 2单克隆抗体试剂套盒行免疫组织化学染色 ,并以染色细胞数目的多少及染色程度为分级标准 ,认为 2 以上 (含 2 )为过表达 (报告为 ～ 的 ,记为阴性 )。结果　该蛋白在A、B期肿瘤中的过表达率为 11 8% (2 /17) ;C、D期肿瘤中过表达率为 5 4 5 % (6 /11)。该蛋白的过表达率在有转移和无转移的病例之间差异有显著意义 (P <0 0 5 )。结论　虽然在结、直肠恶性肿瘤病例中HER 2 /neu的过表达率不高 ,但仍提示HER 2 /neu过表达程度与肿瘤的进展或侵袭性生物学行为相关。     

DNA, RNA and immunohistochemical characterization of the HER-2/neu oncogene in transitional cell carcinoma of the bladder

HER-2/neu overexpression appears to play a role in determining the malignant potential of some human cancers. To date, no urothelial malignancies appear to have been evaluated for HER-2/neu DNA amplification, mRNA expression and protein overproduction. By Southern hybridization we detected DNA amplification and a possible structural rearrangement of the HER-2/neu oncogene in one of 12 bladder tumors. A 14 kb DNA fragment in addition to the expected 12.5 Kb fragment was found. Additionally, the HER-2/neu oncogene was amplified sixfold in the tumor compared to placental DNA. Five of 14 (36%) bladder tumors overexpressed HER-2/neu mRNA three to 38-fold compared to normal urothelium. HER-2/neu overexpression occurred in superficial and invasive tumors. Immunohistochemical analysis was performed on the one tumor with DNA amplification and the 14 tumors evaluated for mRNA expression. The tumor with DNA amplification and three of the five tumors with HER-2/neu mRNA overexpression stained positively for the p185HER-2/neu protein. These findings suggest that DNA amplification occurs infrequently in bladder cancer. Thirty-six percent of bladder cancers overexpress HER-2/neu mRNA. Immunohistochemical analysis with a p185HER-2/neu polyclonal antibody, on formalin fixed, paraffin embedded tissue, was specific for HER-2/neu overexpression but not as sensitive as Northern analysis.     

醛酮还原酶1-B10在胃癌组织中的表达及其临床意义

目的探讨醛酮还原酶1-BIO在胃癌组织中的表达及其与临床病理特征和预后的关系。方法通过荧光定量PCR法检测36例胃癌组织及其配对癌旁组织中AKRlBl0mRNA的表达量,采用免疫组织化学法检测100例原发胃癌组织及70例非肿瘤胃黏膜组织中AKRlB10蛋白表达情况。结果PCR结果显示,91．7％（33／36）癌旁非肿瘤胃黏膜组织中AKRlB10mRNA的表达高于其配对胃癌组织[8．3％（3／36）,P=0．000]。100例胃癌组织中33例（33．0％）AKRIBl0蛋白阳性表达,70例非肿瘤胃黏膜上皮中65例（92．9％）AKRlB10的阳性表达。两者比较,差异有统计学意义（Jp=0．000）。胃癌组织中AKRIBIO的表达与患者性别、年龄、肿瘤部位及分化程度无关（p〉0．05）,而与肿瘤大小（P=O．000）、浸润深度（P=0．004）、淋巴结转移（P=0．028）、远处转移（P=0．031）和临床TNM分期（P=0．000）有关。AKRlB10阳性表达患者的5年生存率显著高于阴性表达者（60．6％比32．8％,P〈0．01）。结论胃癌组织中AKRlB10的表达下调与胃癌进展有关,提示预后不良。     

Role of coordinated molecular alterations in the development of androgen-independent prostate cancer: an in vitro model that corroborates clinical observations

OBJECTIVE: To investigate the role of potential downstream targets of HER-2/neu, including the cell-cycle regulator p27, proliferation-associated protein Ki-67, apoptosis inhibitor Bcl-2, and signal-transduction molecule Akt (which is associated with cell survival), as the development of androgen-independent prostate cancer (AIPC) in patients who are initially responsive to androgen-ablation therapy (AAT) is a significant clinical problem. PATIENTS AND METHODS: Earlier studies showed that high levels of HER-2/neu tyrosine kinase receptor expression as assessed by immunohistochemistry were significantly associated with the development of AIPC, and we hypothesised that HER-2/neu overexpression provides an alternative proliferative stimulus upon androgen depletion. We established a unique clinical model system, comprising patients who received no AAT, or who had preoperative AAT, or those with advanced tumours resistant to AAT. To test our hypothesis in vitro, we stably transfected full-length HER-2/neu cDNA in androgen-responsive LNCaP cells and examined the effects of HER-2/neu overexpression on cell proliferation, apoptosis, androgen-receptor activation, and Akt phosphorylation upon androgen deprivation by using immunohistochemistry and Western blot technique. RESULTS: p27 expression was initially induced on exposure to AAT, and significantly decreased in AIPC (P < 0.001). There was also a significant increase in the Ki-67 index in AIPC (P = 0.001). Elevated Bcl-2 expression was closely associated with AAT (P = 0.002), suggesting that Bcl-2 expression is induced on initial exposure to AAT. Further, Bcl-2 expression was highest in hormone-resistant cancers (P < 0.001). Using the HER-2/neu transfected cell-line model, we confirmed the mechanistic basis of the clinical observations which elucidate the pathway leading to HER-2/neu-mediated androgen independence. On androgen deprivation, the HER-2/neu transfected cells had higher proliferation rates, lower G1 arrest, inhibited p27 up-regulation, a lower apoptotic index, and higher Bcl-2, prostate-specific antigen and phosphorylated Akt expression than the mock-transfected LNCaP cells. CONCLUSION: This study suggests that prostate cancer cells undergo a series of coordinated changes after exposure to AAT, which eventually result in the development of androgen independence. Further, in support of previous results, it appears that a major factor in this process is the induction of HER-2/neu overexpression, which occurs after initial exposure to AAT. HER-2/neu may contribute to the development of androgen independence through: (i) maintaining cell proliferation; (ii) inhibiting apoptosis; and/or (iii) inducing AR activation in a ligand-independent fashion. These effects may be mediated, at least in part, through activation of the PI3K/Akt pathway.