CK20和Ki-67在膀胱移行细胞癌中的表达及临床意义

目的探讨细胞角质蛋白20（CK20）和细胞核相关抗原（Ki-67）在膀胱移行细胞癌中的表达及临床意义。方法对经临床确诊的膀胱移行细胞癌患者癌组织标本中CK20（66例）和Ki-67（55例）的表达结果进行回顾性分析。结果CK20在膀胱移行细胞癌表达的阳性率为68．2％（45／66）。CK20表达的阳性指数与膀胱移行细胞癌临床分期和细胞分级均呈正相关（P〈0．05）。复发膀胱移行细胞癌组CK20阳性表达率92．3％（12／13）高于初发膀胱移行细胞癌组62．3％（33／53）（P〈0.05）。Ki-67在膀胱移行细胞癌表达的阳性率为63．6％（35／55）。Ki-67表达的阳性指数与膀胱移行细胞癌细胞分级呈正相关（P〈0.05）。复发膀胱移行细胞癌组Ki-67阳性表达率90．9％（10／11）高于初发膀胱移行细胞癌组56．8％（25／44）（P〈0．05）。Ki-67和CK20的表达阳性指数呈正相关（r=0．493,P〈0．05）。结论研究结果表明CK20和Ki-67的表达与膀胱移行细胞癌的生物学行为相关,因此CK20和Ki-67可作为膀胱移行细胞癌肿瘤病理分级分期以及预后判断的辅助手段。     

膀胱移行细胞癌中凋亡相关蛋白survivin及caspase3的表达及相关性

目的探讨凋亡相关蛋白survivin及caspase3的表达与膀胱移行细胞癌发生及发展的关系。方法应用SP免疫组织化学法检测69例膀胱移行细胞癌石蜡切片,应用免疫印迹法（Western blotting）检测33例膀胱移行细胞癌新鲜组织中survivin和caspase3表达的情况,结合临床资料进行分析。结果免疫组化结果显示,survivin在膀胱移行细胞癌标本中的表达阳性率为88．4％（61／69）,而正常对照组均呈阴性;caspase3在膀胱移行细胞癌标本中的表达阳性率为59．4％（41／69）,但是与对照组相比无明显差异。Western blotting结果显示,93．9％（31／33）的肿瘤组织可见survivin蛋白表达,而对照组全部阴性表达;81．8％（27／33）的肿瘤标本可见caspase3蛋白表达,对照组中阳性表达率为80％（8／10）,实验组与对照组间无明显差异。在Ⅲ级膀胱移行细胞癌中survivin的表达强度较Ⅰ级为高,二者之间差异有显著性（P〈0．05）。survivin和caspase3的表达与肿瘤的临床分期无关（P〉0．05）,而且survivin的表达与easpase3的表达之间也没有相关性。结论survivin蛋白在膀胱移行细胞癌中特异性表达,其表达的上调可能提示肿瘤分化不良。Caspase3蛋白的表达情况与膀胱移行细胞癌关系不密切。     

凋亡相关蛋白Survivin及Caspase-3在膀胱移行细胞癌中表达及临床意义

目的：探讨凋亡相关蛋白Survivin及Caspase-3在膀胱移形细胞癌表达及其临床意义。方法：应用SP免疫组织化学法检测45例膀胱移行细胞癌及10例正常膀胱黏膜组织石蜡切片中Survivin和Caspase-3表达的情况,结合临床资料进行分析。结果：Survivin在膀胱移形细胞癌标本中的表达阳性率为68．9%（31/15）．而正常对照组中无一例呈阳性表达;Caspase-3在膀胱移行细胞癌标本中的表达阳性率为37．8%（17/45）．与对照组阳性率90％（9／10）相比差异有统计学意义（P〈O．05）。Survivin的表达与膀胱移行细胞癌的组织学分级、初发和复发显著相关（P〈O．05）,但与临床病理分期、肿瘤数日无关;Caspase-3的表达与膀胱移行细胞确的初发复发相关,但与组织学分级、肿瘤数目、临床分期均无关。相关性分析表明,膀胱移行细胞癌中Survivin的表达与Caspase-3表达呈负相关。结论：Survivin在膀胱癌组织中选择性表达与膀胱移行细胞癌的分化程度密切相关．Caspase-3蛋白在膀胱移行细胞癌中表达下降,Survivin及Caspase-3蛋白对于判断膀胱移行细胞确预后有重要临床指导意义。     

O6-甲基鸟嘌呤-DNA甲基转移酶在膀胱移行细胞癌中的表达及临床意义

目的：研究O^6-甲基鸟嘌呤-DNA甲基转移酶（MGMT）在膀胱移行细胞癌（TCCB）组织中的表达及其与临床病理特征的关系。方法：应用免疫组织化学SP法检测60例膀胱移行细胞癌中MGMT的表达,统计分析其与肿瘤临床病理的关系。结果：膀胱癌组织中MGMT阳性表达率为35．0％（21／60）,明显低于癌旁组织（76．7％．46／60）及正常膀胱组织（86．7％,13／15）（P〈0．015;MGMT蛋白的表达与膀胱移行细胞癌组织分化程度有关（P＜0．05）。而与临床分期无明显关系。结论：MGMT在膀胱移行细胞癌的发生和发展中起抑制作用;MGMT蛋白的表达可能是预断膀胱癌预后的重要指标。     

膀胱癌错配修复基因hMSH2表达的研究

目的 探讨错配修复基因hMSH2在膀胱移行细胞癌中的表达情况及其与肿瘤细胞增殖、凋亡的关系.方法 采用免疫组化法检测101例膀胱移行细胞癌错配修复基因hMSH2以及细胞增殖抗原Ki-67的表达情况,原位末端标记(TUNEL)法检测肿瘤细胞凋亡情况.结果 hMSH2表达于非膀胱肿瘤尿路上皮及部分膀胱移行细胞癌的细胞核,膀胱移行细胞癌组低表达率较非膀胱肿瘤膀胱组织组高(P=0.004,＜0.05);膀胱移行细胞癌pT2-pT4组中hMSH2的低表达率比pTis-pT1组的低表达率高(P=0.016,＜0.05);G1、G2、G3三组间低表达率的总体差别有统计学意义(P=0.033,＜0.05),G1+G2组与G3组之间差别有统计学意义(P=0.036＜0.05);有无淋巴结转移组差别无统计学意义(P=0.317).hMSH2弱表达组与强表达组间凋亡指数(AI)均值差异非常显著(P＜0.01),Ki-67标记指数(Ki-67 LI)均值差异不显著(P＞0.05),AI/KI值差异经统计学分析具有统计学意义(P＜0.05).结论 hMSH2基因突变或功能缺失与膀胱移行细胞癌的发生有关,可能系肿瘤发生过程中的重要事件,并可能与肿瘤细胞凋亡有关,而与增殖活性无关.     

凋亡相关基因Survivin及Caspase-3 mRNA在膀胱移行细胞癌中的表达及其意义

目的探讨凋亡相关基因survivin及Caspase-3 mRNA的表达与膀胱移行细胞癌发生及发展的关系。方法应用逆转录-聚合酶链式反应（RT-PCR）检测33例膀胱移行细胞癌组织中survivin和Caspase-3 mRNA表达的情况，结合临床资料进行分析。结果93．9％（31／33）的肿瘤组织可检测到survivin mRNA表达，而对照组全部阴性表达，二者有统计学意义；81．8％（27／33）的肿瘤标本可检出Caspase-3 mRNA，而对照组中检出率为80％（8／10），实验组与对照组间无明显差异。在Ⅲ级膀胱移行细胞癌中survivin mRNA的表达强度较Ⅰ级为高，二者间有统计学意义（P〈0．05）。survivin和Caspase-3的表达与肿瘤的临床分期无关（P〉0．05）。结论survivin mRNA在膀胱移行细胞癌中有特异性表达，其高表达提示肿瘤分化不良。阻断suvivin mRNA的表达可能为膀胱肿瘤的治疗提供新的途径。     

膀胱移行细胞癌Survivin表达临床特征分析

目的：探讨Survivin蛋白在不同病理类型及不同临床分期的膀胱移行细胞癌中的表达规律。方法：膀胱癌组为膀胱移行细胞癌44例肿瘤组织保存腊块,其中病理分级I级16例、Ⅱ级16例、Ⅲ级12例;临床分期表浅型24例、浸润型20例;另取10例非肿瘤患者的正常膀胱黏膜做对照。采用免疫组织化学SABC 进行Survivin表达的定位和半定量检测。结果：正常对照组Survivin阳性表达率为0％（O／10）,44例膀胱移行细胞痛中34例Survivin蛋白表达阳性,阳性率为77.3％（34／44）,显著高于正常对照组。分项统计中,肿瘤病理分级I级组（56.3％,9／16）、Ⅱ级组（87.5％,14／16）、Ⅲ级组（91.7％．11／12）及表浅型（79.2％．19/24）、浸润刚（75.0％,15／20）Survivin表达阳性率均显著高于正常对照组。相关分析表明,Survivin表达与肿瘤病理分级密切相关（x^2=6.395,P〈0.05）,与临床分期无显著相关关系（x^2=0.108．P〉0.05）。结论：Survivin异常表达在膀胱移行细胞癌的发生发展中起重要作用,Survivin表达检测可成为膀胱移行细胞癌预后指标之一。     

细胞核增殖抗原与E-钙黏蛋白在膀胱移行细胞癌中的表达及临床意义

目的 探讨细胞核增殖抗原(Ki-67)、E-钙黏蛋白(E-cadherin)在膀胱移行细胞癌中的表达及其临床意义.方法 采用免疫组织化学方法检测Ki-67、E-cadherin在48例膀胱移行细胞癌组织中的表达,统计分析其表达与膀胱移行细胞癌临床病理、分期之间的关系.结果 Ki-67、E-cadherin在膀胱移行细胞癌中的阳性表达率分别是83.3％和47.9％,两者呈负相关(P＜0.05).两者表达水平与肿瘤临床分期及病理分级明显相关(P＜0.05).结论 检测Ki-67与E-cadherin有助于判断膀胱移行细胞癌侵袭和转移能力.     

survivin和环氧化酶-2在膀胱移行细胞癌组织中的表达及其相关性

目的 探讨膀胱移行细胞癌组织中凋亡抑制蛋白survivin和环氧化酶-2（cox-2）的表达与肿瘤生物学行为之间的关系及其二者的相关性。方法应用免疫组化Envision法，检测42例膀胱移行细胞癌组织标本和10例非肿瘤正常膀胱组织（正常对照组）中survivin和cox-2的表达。结杲42例膀胱移行细胞癌组织中survivin和COX-2表达的阳性率分别为78．6％和81．0％，正常对照组中均未见表达；survivin表达强度与肿瘤分期及复发呈正相关关系（P〈O．05），而与肿瘤分级无关（P〉O．05）；cox-2表达强度与肿瘤分级和分期呈正相关关系（P〈0．05），而与肿瘤复发无关（P〉0．05）；survivin和cox-2在膀胱移行细胞癌中的表达密切相关（rs=0．327，P〈0．05）。二者的表达与患者的性别、年龄、术时肿瘤的大小、数目及部位无关（P〉0．05）。结论Survivin和cox-2在膀胱移行细胞癌组织中普遍表达，且表达密切相关，二者可能在膀胱癌的发生、发展过程中起重要作用，并与浸润关系密切。     

Clusterin蛋白在肾癌组织中的表达及临床意义

目的 探讨clusterin蛋白在肾癌组织中的表达及临床意义。方法 制作87例肾癌组织芯片，应用免疫组织化学方法检测肾癌组织中clusterin蛋白表达，应用TUNEL方法检测细胞凋亡情况，分析clusterin蛋白表达与肾癌细胞凋亡及临床病理学参数之间的相关性。结果 68例有效检测的肾癌组织标本中，clusterin蛋白过度表达15例（22％），且其表达与肿瘤浸润有显著相关性（P=0．004），71％（5／7）的T3肿瘤clusterin蛋白呈过度表达，84％（51／61）的T1、T2肿瘤clusterin蛋白为正常表达。57％（4／7）伴有淋巴结转移的肾癌组织标本clusterin蛋白呈过度表达，阳性率高于无淋巴结转移者（18％，11／61，P=0．059）。clusterin蛋白表达与肿瘤凋亡指数（AI）呈负相关（P=0．0138），61％（8／13）clusterin过度表达的肿瘤表现为低水平的AI值，78％（39／50）clusterin正常表达的肿瘤AI值为高或中水平。结论 肾癌组织中clusterin蛋白过度表达可能与其抗凋亡作用有关，与肾癌的恶性临床表型相关，可作为判断肾癌恶性程度的指标之一。     

Livin在膀胱移行细胞癌中的表达及其与ki-67的关系

目的研究凋亡抑制蛋白家族（IAP家族）新的凋亡抑制因子livin在膀胱移行细胞癌（BTCC）中的表达及其与肿瘤分级、分期的关系及livin和ki-67在BTCC中表达的关系。方法采用免疫组化SP法对36例BTCC和8例正常膀胱黏膜livin和ki-67的表达进行检测,分析两者在膀胱癌组织和非膀胱癌组织中的表达,livin的表达与BTCC病理学分级、临床分期及与ki-67表达的关系。结果Livin在8例正常膀胱组织中均不表达,而在36例BTCC组织中的阳性表达率为86．1％（P〈0．01）。livin的表达和BTCC的病理分级,临床分期、k-67的表达无明显相关（P〉0．05）。结论细胞凋亡抑制因子livin在BTCC组织中表达上调,提示livin可能通过抑制细胞凋亡,对BTCC的发生发展起重要作用;但是我们的实验表明livin与调控细胞周期,促进细胞增殖无明显相关。Livin在膀胱癌中的高表达,有望成为一种有效、敏感的瘤标,并为BTCC的基因治疗提供新靶点。     

Expression of the secreted form of clusterin protein in renal cell carcinoma as a predictor of disease extension

OBJECTIVES: To evaluate the significance of clusterin expression in surgically resected renal cell carcinoma (RCC) specimens. PATIENTS AND METHODS: Normal kidney and RCC specimens were obtained from 131 patients who had radical surgery. The expression of clusterin protein was analysed by immunohistochemical staining with an antibody recognizing all isoforms of clusterin. Cell proliferative activities and apoptotic features in these specimens were investigated using Ki-67 immunostaining and the terminal deoxynucleotidyl transferase mediated dUTP nick-end labelling assay, respectively. Findings were evaluated in relation to several clinicopathological factors. RESULTS: There were various levels of clusterin expression in 128 of the 131 RCC specimens, while 37 of 131 normal kidney tissues (28.2%) had no clusterin staining. Clusterin protein was present in the cytoplasm of both normal and cancer cells, but there was no nuclear staining identified in either type of cell. The expression level of clusterin protein in RCC tissues was significantly related to tumour stage and grade, but not to age, gender or histological cell type. Cell proliferative activity in RCC specimens was significantly associated with clusterin expression, while the apoptotic index was inversely related to clusterin expression. Furthermore, recurrence-free survival in patients with strong clusterin expression was significantly lower than that in those with weak expression. CONCLUSIONS: These findings suggest that the secreted form of clusterin may be involved in the progression of RCC, and that overexpression of clusterin could be a useful prognostic variable after radical surgery in patients with RCC.     

人类胰腺癌Ki—67基因表达的研究

目的 应用原位杂交结合免疫组化分析胰腺癌Ｋｉ－６７基因的ｍＲＮＡ转录和蛋白翻译,研究该基因结构和功能表达的关系。方法 胰腺癌４０例,正常胰腺及良性病变９例;扁桃体组织及Ｈｅｌａ细胞作为阳性对照。通过地高辛标记的Ｋｉ－６７ｃＲＮＡ探针原位杂交和Ｋｉ－６７等效单克隆抗体的免疫组化分析该基因的ｍＲＮＡ转录和蛋白翻译。结果正常胰腺及良性病变Ｋｉ－６７ 指数均小于２０％;胰腺主分化腺癌、低分化腺癌各２０例,     

cyclinD1在膀胱移行细胞癌中的表达及与肿瘤增殖活性的关系

目的：研究cyclinD1在膀胱移行细胞癌（TCC）标本中的表达情况及其与肿瘤增殖活性的关系。方法：应用免疫组织化学SP法检测45例TCC和12例正常膀胱cyclinD1和Ki-67抗原的表达。结果：（1）正常膀胱组织无cyclinD1表达，Ki-67抗原的表达低于TCC（P=0.011)。（2）随着肿瘤分期。分级的增高，cyclinD1阳性率下降（P=0.014,P=0.034);(3)cyclinD1和Ki-67抗原的表达呈负相关（r=-0.4109,P=0.005)。结论：cyclinD1在TCC的早期起重要作用；Ki-67指数能准确地评估TCC的生物学行为，二者可作为TCC有重要意义的肿瘤标志物。     

CD147 mRNA及蛋白在膀胱移行细胞癌中的表达及临床意义

目的：研究膀胱移行细胞癌（BTCC）及正常膀胱组织中CD147mRNA及其蛋白的表达,探讨其临床意义。方法：采用原位核酸分子杂交和免疫组化S-P法检测67例BTCC组织和12例正常膀胱组织中CD147mRNA和蛋白的表达情况,探讨其与BTCC部分临床生物学特性的相关性,及CD147mRNA和蛋白表达的一致性。结果：正常膀胱组织未见CD147mRNA和蛋白表达。BTCC组织中CD147mRNA及蛋白阳性率分别为59．7％和68．7％。CD147蛋白表达与病理分级显著相关（r=0．482,P〈0．001）,与临床分期朱见相关;CD147mRNA表达定位于胞浆,与膀胱癌病理分级、临床分期未见相关;CD147mRNA和蛋白表达呈正相关（r=0．285,P=0．019）。结论：BTCC中有CD147mRNA和蛋白的表达,其表达呈正相关,CD147有可能成为判断BTCC侵袭力的重要指标,阻断CD147的表达可能成为治疗膀胱肿瘤的新靶点。     

膀胱癌组织端粒酶活性的研究

目的　探讨膀胱癌组织端粒酶活性的临床意义。　方法　应用银染端粒重复序列扩增法检测 42例膀胱癌及其癌旁组织的端粒酶活性。　结果　 3例正常膀胱组织端粒酶表达均阴性 ;42例膀胱癌组织中端粒酶表达阳性 35例 (83 .3 % ) ,癌旁组织端粒酶表达阳性 7例 (16 .7% ) ;浸润癌或有淋巴结或远处转移者端粒酶表达阳性率高于非浸润癌或无转移者 ,但差别无显著性意义 (P >0 .0 5 )。　结论　端粒酶是膀胱癌较理想的肿瘤标记物之一。     

Cellular apoptosis, proliferation and bcl-2 Bax expression in colorectal carcinoma and their association with tumor prognosis

OBJECTIVE: To investigate the relationship between cellular apoptosis, proliferation and bcl-2/bax expression in the tissue of colorectal carcinoma and biological behavior of the tumor. METHOD: The apoptotic cells index (AI) in situ were identified by the terminal deoxynucleotidyl transfer-mediated dUTP-biotin nick end labeling (TUNEL) and immunohistologic staining for Ki-67 proliferation index (KI), bcl-2/bax protein expression was performed on archival material from 77 adenocarcinomas. RESULT: The mean AI and KI were 5.3/45.4 in early stage cancer, 5.6/48.7 in non-metastatic cancer, 6.6/55.3 in advanced cancer and 8.9/60.1 in cancer with distant metastasis respectively. There was a positive relationship between the AI and KI in each specimen. bcl-2 and bax was expressed as 54.5%, 74% in carcinoma and highly differentiated carcinoma. In the univariate analysis, significantly longer survival time was observed in the subgroup of bcl-2 positive carcinoma with low AI. In the multivariate analysis, tumor stage, tumor type, KI, and bcl-2 expression were independent risk factors for prognosis. CONCLUSION: The data indicate that abnormally exchanged AI/KI, bcl-2/bax expression appears to be an event associated with tumor progression and apoptosis might also reflect the proliferative activity of human colorectal carcinoma.     

Immunohistochemical study of p53 and Ki-67 overexpression in grade 3 superficial bladder tumor in relationship to tumor recurrence and prognosis]

PURPOSE: The major drawback of the current treatment for superficial bladder tumor is the high rate of recurrence. Especially, the tumor with grade 3 component has a tendency to recur and progress in stage. However, we have difficulty in predicting tumor recurrence and stage progression accurately by conventional clinicopathological factors. We evaluated the efficacy of p53 and Ki-67 overexpression as a predictor of recurrence or prognosis in patients with superficial bladder tumor of grade 3. MATERIALS AND METHODS: Samples were obtained from 41 patients with superficial transitional cell carcinoma of the bladder of grade 3 who were treated by transurethral resection (TUR). The immunohistochemical study was performed using the antibodies against the p53 protein and Ki-67 antigen on formalin-fixed, paraffinembedded tissue specimens from initial tumors. We evaluated the correlation between these results and several clinicopathological factors. RESULTS: The p53 index and the Ki-67 index in pTa, pT1a and pT1b tumors were 26.4 +/- 30.1%, 28.6 +/- 30.0%, and 34.6 +/- 32.6% (p53) and 20.5 +/- 22.5%, 20.0 +/- 29.3%, and 29.2 +/- 28.4% (Ki-67). There was no significant difference between the each index and tumor stage. Eighteen cases (43.9%) had intravesical recurrence. The p53 index of the initial tumor from the tumor free cases (n = 23), recurrent cases without stage progression (n = 12), and stage progression cases (n = 6) were 19.7 +/- 28.2%, 42.0 +/- 28.7%, and 42.5 +/- 32.0%. Between the recurrence-free cases and the recurrent cases without progression, the p53 index of the initial tumor had statistical significance (p < 0.05). The Ki-67 index was shown to be the same pattern as the p53 index, but there was not statistical significance. Four of patients with stage progression had tumor progression within six months. Three of the patients with tumors with stage progression died of the cancer. In multivariate analysis, tumor multiplicity (p = 0.01), BCG intravesical instillation (p = 0.04), p53 index (p = 0.01) and Ki-67 index (p = 0.02) were the positive risk factors for tumor recurrence, but only the p53 index was the positive risk factor for prognosis fo the patients (p = 0.03). CONCLUSION: These results suggest that the immunohistochemical study of p53 overexpression is a useful predictor for tumor recurrence and prognosis in patients with superficial bladder tumor with grade 3.     

Tissue microarray sampling strategy for prostate cancer biomarker analysis

High-density tissue microarrays (TMA) are useful for profiling protein expression in a large number of samples but their use for clinical biomarker studies may be limited in heterogeneous tumors like prostate cancer. In this study, the optimization and validation of a tumor sampling strategy for a prostate cancer outcomes TMA is performed. Prostate cancer proliferation determined by Ki-67 immunohistochemistry was tested. Ten replicate measurements of proliferation using digital image analysis (CAS200, Bacus Labs, Lombard, IL, USA) were made on 10 regions of prostate cancer from a standard glass slide. Five matching tissue microarray sample cores (0.6 mm diameter) were sampled from each of the 10 regions in the parallel study. A bootstrap resampling analysis was used to statistically simulate all possible permutations of TMA sample number per region or sample. Statistical analysis compared TMA samples with Ki-67 expression in standard pathology immunohistochemistry slides. The optimal sampling for TMA cores was reached at 3 as fewer TMA samples significantly increased Ki-67 variability and a larger number did not significantly improve accuracy. To validate these results, a prostate cancer outcomes tissue microarray containing 10 replicate tumor samples from 88 cases was constructed. Similar to the initial study, 1 to 10 randomly selected cores were used to evaluate the Ki-67 expression for each case, computing the 90th percentile of the expression from all samples used in each model. Using this value, a Cox proportional hazards analysis was performed to determine predictors of time until prostate-specific antigen (PSA) recurrence after radical prostatectomy for clinically localized prostate cancer. Examination of multiple models demonstrated that 4 cores was optimal. Using a model with 4 cores, a Cox regression model demonstrated that Ki-67 expression, preoperative PSA, and surgical margin status predicted time to PSA recurrence with hazard ratios of 1.49 (95% confidence interval [CI] 1.01-2.20, p = 0.047), 2.36 (95% CI 1.15-4.85, p = 0.020), and 9.04 (95% CI 2.42-33.81, p = 0.001), respectively. Models with 3 cores to determine Ki-67 expression were also found to predict outcome. In summary, 3 cores were required to optimally represent Ki-67 expression with respect to the standard tumor slide. Three to 4 cores gave the optimal predictive value in a prostate cancer outcomes array. Sampling strategies with fewer than 3 cores may not accurately represent tumor protein expression. Conversely, more than 4 cores will not add significant information. This prostate cancer outcomes array should be useful in evaluating other putative prostate cancer biomarkers.