【作者】 木尔扯尔；

【导师】 文彬；

【作者基本信息】 川北医学院 ， 人体解剖与组织胚胎学， 2012， 硕士

【摘要】 目的：通过检测β-catenin、wisp-1和caspase9蛋白、基因在结肠癌中的表达，分析3个因子与结肠肿瘤临床病理学特征的关系，探讨其在结肠癌发生中的作用。方法：运用免疫组化Evision法检测β-catenin、wisp-1和caspase9蛋白在138例外检结肠腺癌、37例结肠腺瘤和66例结肠切缘中的表达；运用实时荧光定量RT-PCR法检测β-catenin、wisp-1和caspase9 mRNA在37例结肠肿瘤及其结肠切缘组织中的表达；实验结果结合临床病理资料进行统计分析。结果：1.β-catenin蛋白在结肠腺瘤、癌旁组织及结肠癌中阳性表达高于结肠切缘组织(p<0.05)；wisp-1蛋白在结肠癌、癌旁组织阳性表达高于切缘组织（p<0.05）；β-catenin蛋白、wisp-1蛋白在结肠癌中的≥++阳性表达高于结肠腺瘤和癌旁组织（p<0.05）。Caspase9蛋白在结肠切缘、结肠腺瘤、癌旁组织及结肠癌中的阳性表达有依次降低的趋势，但相互进行比较无统计学意义（p>0.05），各组间≥++阳性率比较亦无统计学意义（p>0.05）。2.β-catenin蛋白的阳性表达与淋巴结转移、临床分期以及浸润深度有关(p<0.05)，与患者的性别、年龄、肿瘤类型、分化程度无明显关系(p>0.05)。wisp-1蛋白阳性表达与淋巴结转移及组织浸润深度有关（p <0.05），与患者性别、年龄、肿瘤类型、分化程度、duke分期及肿瘤部位无关（p>0.05）。caspase9蛋白阳性表达率与临床各项指标之间均无统计学意义（p>0.05）。3.β-catenin、wisp-1、caspase9 mRNA在结肠癌中平均表达水平分别是正常结肠中表达的1.946倍(p<0.05)，1.349倍（p>0.05）和1.587倍（p>0.05）。β-catenin mRNA表达水平与结肠癌的部位、肿瘤类型和淋巴结转移有关(p<0.05)，与患者性别、年龄、肿瘤分化程度以及浸润深度无关（p>0.05）；wisp-1、caspase9 mRNA表达水平与肿瘤类型有关，与患者性别、年龄、肿瘤部位、分化程度以及是否转移、浸润深度均无关（p>0.05）。4.β-catenin与wisp-1蛋白表达的相关系数为0.423（p<0.001），基因表达的相关系数为0.461（p<0.005）；wisp-1与caspase9蛋白表达之间相关系数为0.183（p<0.05），基因表达之间的相关系数为0.526（p<0.001）；β-catenin与caspase9在蛋白表达水平上的相关系数为0.110（p>0.05），基因水平的相关系数为0.094（p>0.05）。结论：1.β-catenin蛋白和基因在结肠癌中高表达，结合其临床病理特点，提示该因子可能与结肠肿瘤的发生发展及预后有关。2.wisp-1蛋白在结肠癌中高表达，显著高于结肠腺瘤及正常粘膜组织，且与肿瘤的浸润深度、淋巴结转移关系密切，提示其可能与肿瘤的发生发展及临床预后有关；但wisp-1 mRNA在结肠癌和正常结肠组织中的表达无显著差异，提示wisp-1 mRNA在结肠癌中的作用尚不明确。3.Caspase9蛋白及基因在结肠癌、结肠腺瘤、正常结肠组织中的表达均无显著差异，提示其在结肠癌中的作用尚不明确。4.β-catenin与wisp-1蛋白、基因表达均呈正相关关系，提示两因子在结肠癌发生发展中可能单独或共同促进肿瘤的发生。更多还原

【Abstract】 Objective：To explore the gene，protein expression ofβ-catenin、wisp-1、caspase-9 in Colon tumor and analyze the relationship between theexpression and clinical pathology of Colon tumor，correlations between theβ-catenin、wisp-1、caspase-9 protein and gene.Methods：The protein expression ofβ-catenin、wisp-1、caspase-9 weredetected by immunohistochemistry（IHC）in 138 cases of primary coloncancer tissue、37 cases of colon adenoma tissue and 66 cases of normal colonmucosa tissue；the mRNA expression ofβ-catenin、wisp-1、caspase-9 weredetected by real-time fluorescence quantitative PCR（RT-PCR）in 37 cases offresh primary colon tumor and 37 cases of fresh normal colonic mucosa；Theexperimental results combined were statistically analyzed with the clinicalpathological sample data.Results：1. All the statistics of the positive expressions ofβ-cateninprotein in colon cancer、colon adenoma and the tissue of adjacent coloncancer were higher than that in normal colon mucosa (p<0.05)；The positiveexpressions of wisp-1 protein in both the colon cancer and the tissue ofadjacent colon cancer were higher than normal colon mucosaexpression(p<0.05); The strong positive(≥++)of bothβ-catenin and wisp-1protein in Colon cancer was higher than that in colon adenoma and the tissueof adjacent colon cancer (p<0.05). The positive expression rate of caspase9 protein in colon cancer，colon adenoma，tissue of adjacent colon cancer andnomal colon tissue shows a rise trend，but comparing the expression rate ofeach group，there is no statistically significant difference (p>0.05)；comparingthe strong expression rate(≥++) of each group no statistically significantdifference either (p>0.05). 2.β-catenin protein expression had closerelationship with clinical stages、infiltration depth and lymph nodemetastasis(P<0.05)，there is no significant correlation betweenβ-cateninprotein positive expression and sample’s gender，age，tumor types anddifferentiation degree (p>0.05).wisp-1 protein expression just had closerelationship with infiltration depth and lymph node metastasis(P<0.05)，butno significant correlation between wisp-1 positive expression rate andsamples’gender，age，tumor types，differentiation degree and tumor location(p>0.05)；Caspase9 protein positive expression rate has no significantcorrelation with each clinical index. 3.In colon cancer，although the averageexpression ofβ-catenin、wisp-1、caspase9 mRNA were respectively1.946，1.349 and 1.587 times the level in matched normal colon tissue，butonly the result ofβ-catenin mRNA difference had Statistics Meaning(P<0.05).β-catenin mRNA expression had close relationship with tumorlocation、tumor types and lymph node metastasis(P<0.05)，yet had norelationship with gender、age、infiltration depth and differentiation degree(p>0.05).wisp-1 and caspase9 mRNA expression only had relationship withtumor types and no significate relationship with any other of pathologicalfeatures.4.There were positive correlations betweenβ-catenin protein andwisp-1 protein（r=0.423，p < 0.001），β-catenin mRNA and wisp-1 mRNA (r=0.461，p<0.005)，Wisp-1 protein and caspase9 protein（r =0.183，p<0.05），Wisp-1 mRNA and caspase9 mRNA（r =0.526，p<0.001），respectively. There were no correlations betweenβ-catenin protein andcaspase9 protein（r =0.110，p > 0.05），β-catenin mRNA and caspase9mRNA（r =0.094，p > 0.05）.Conclusions：1. Both ofβ-catenin protein and mRNA show highexpressions，which with their clinical features suggests thatβ-catenin may becorrelated with the development and diagnosis of colon tumor. 2. The wisp-1protein in colon tumor shows high expression which is obviously higher thanthat of Colonic adenoma and normal mucosa tissue and closely correlated totumor infiltration depth，lymph node metastasis. This suggests that thewisp-1 protein may be correlated with the development and diagnosis ofcolon tumor. However，there is no significant difference between theexpressions of wisp-1 mRNA in colon cancer and the normal colonic tissue，which indicates that the role of wisp-1 mRNA is not clear in the developmentof colon tumor.3.This is no significant difference in protein and mRNAexpression levels of caspase9 in colon cancer，colon adenoma and normalcolonic mucosa，which suggests the role of caspase-9 in the development ofcolon cancer was not clear either.4. The protein and mRNA expressions ofβ-catenin and wisp-1 have a positive correlation，which suggests thatβ-catenin and wisp-1 are involved in the development of colon cancer，eitherjointly or independently.更多还原