【作者】 易晓佳；

【导师】 章宗籍； 申丽娟； 张华献； 钱忠义；

【作者基本信息】 昆明医学院 ， 病理学与病理生理学， 2008， 硕士

【摘要】 目的:研究P57^kip2基因在人肝细胞癌（Hepatocellular carcinoma,HCC）不同阶段的杂合性缺失（Loss of heterozygosity,LOH）、微卫星不稳定性（Microsatellite instability,MSI）及其mRNA和蛋白表达情况,以及它们之间的相互关系,以探讨HCC发生的分子生物学机制。方法:运用PCR—聚丙烯酰胺凝胶电泳—银染法对30例肝癌组织及其癌周肝硬化组织P57^kip2基因所在染色体区域的3个微卫星位点（D11S1760,D11S922和D11S1338）进行LOH和MSI检测,同时运用原位分子杂交和免疫组化技术分别检测30例正常肝组织、30例癌周肝硬化组织和30例肝癌组织中P57^kip2mRNA及P57^kip2蛋白的表达情况。结果:1.LOH及MSI检测:30例肝癌组织中3个位点均未发生LOH:在2个微卫星位点发生MSI,MSI总阳性率为16.7%。2.P57^kip2mRNA表达:正常肝组织未见表达,癌周肝硬化组与肝癌组阳性表达率均为26.7%（8/30）。肝癌组高分化、中分化和低分化三组之间P57^kip2mRNA表达差异有统计学意义。3.P57^kip2蛋白表达:正常肝组织未见表达,癌周肝硬化组与肝癌组阳性表达率均为56.67%（17/30）。肝癌组高分化、中分化和低分化三组之间P57^kip2蛋白表达差异有统计学意义。4.肝癌组微卫星位点D11S1760发生MSI与P57^kip2mRNA和蛋白表达均有关联性。5.肝癌组P57^kip2mRNA与蛋白表达之间有关联性。结论:1.P57^kip2mRNA和蛋白表达异常提示其可能在HCC发生过程中起重要作用。2.与P57^kip2基因同一区域的3个微卫星位点均未发生LOH,提示这3个位点不是P57^kip2基因LOH的频发位点,与P57^kip2mRNA和蛋白表达异常无关。3.P57^kip2基因同一区域的2个微卫星位点发生MSI,同时伴有P57^kip2mRNA和蛋白表达异常,提示MSI可能是肝癌发生过程中P57^kip2mRNA和蛋白表达异常的原因之一。4.微卫星位点D11S1760发生MSI与P57^kip2mRNA和蛋白表达异常有关联,提示该位点可能是肝癌P57^kip2表达异常的特异性标志位点。更多还原

【Abstract】 Objective:To study the loss of hererozygosity（LOH）and microsatellite instability（MSI）of P57^kip2and expressions of P57^kip2mRNA & protein in human hepatocellular carcinoma（HCC）in order to explore the molecular mechanism of hepatocarcinogenesis.Methods:MSI and LOH of three microsatellite locis （D11S1760,D11S922and D11S1338）were detected where the P57^kip2gene is in by using PCR-Polyacrylamide gel electrophoresis-silver staining method in 30 cases of HCC.Immunohistochemistry staining and in situ hybridization（ISH）were also used to show the expression of P57^kip2 protein and mRNA in pericancerous cirrhosis and HCC.Results:1.LOH and MSI detected:LOH was not identified in 30 cases on three microsatellite locies;there were 2 microsatellite locies showing MSI,the total rate of MSI was 16.7%2.Expression of P57^kip2mRNA:There were no expressions in normal liver tissue.Expressions both in pericancerous cirrhosis and hepatocellular carcinoma was 26.67%（8/30）.There was significant difference between the expressions in high differentiated HCC tissues,moderately differentiated HCC tissues and low differentiated HCC tissues.3.Expression of P57^kip2protein:There were no expressions in normal liver tissue.Expressions in precancerous cirrhosis and hepatocellular carcinoma was 56.67%（17/30）respectively.There was no significant difference between the expressions in high differentiated HCC tissues, moderately differentiated HCC tissues and low differentiated HCC tissues.4.There was correlation between the MSI of D11S1760 locies and the expression of P57^kip2mRNA;there was correlation between the MSI of D11S1760 locies and the expression of P57^kip2protein.5.There was positive correlation between expression of P57^kip2mRNA and expression of P57^kip2protein.Conclusion:1.The disorder expressions of P57^kip2mRNA and protein indicated that P57^kip2may be involved in hepatocarcinogenesis and prognosis.2.LOH was not identified on three microsatellite locies,which indicated that there may be another reason for the disorder expression of P57^kiP2.3.Two microsatellite locis showed MSI where the P57^kip2gene is in, with the disorder expression of P57^kip2mRNA and protein,indicated that MSI may be one of the reasons that lead to the disorder expression of P57^kip2mRNA and protein in hepatocarcinogenesis and prognosis.4.There was correlation between the MSI of D11S1760 locies and the expression of P57^kip2mRNA and protein,which indicated that MSI of D11S1760 may the specifical marker for the disorder expression of P57^kip2in HCC.更多还原