【作者】 王翠欣；

【导师】 章宗籍； 申丽娟；

【作者基本信息】 昆明医学院 ， 病理学与病理生理学， 2007， 硕士

【摘要】 目的：研究P57^kip2基因在人体肝细胞癌（Hepatocellular carcinoma，HCC）不同阶段的杂合性缺失（Loss of heterozygosity，LOH）及其mRNA和蛋白表达情况，以及三者之间的关系，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例肝癌组织与P57^kip2基因同一区域的3个微卫星位点（D11S1397，D11S1318和D11S4046）进行LOH的检测，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例癌周肝硬化和30例肝癌组织中P57^kip2mRNA及P57^kip2蛋白的表达情况。结果：1．LOH检测：30例肝癌组织中在3个微卫星位点均未出现LOH。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为26.7％（8／30）。肝癌组分化好和分化差阳性表达率之间差异无统计学意义。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分别为56.67％（17／30）和63.33％（19／30）。肝癌组分化好者与分化差者阳性表达率之间差异无统计学意义。4．肝癌3个微卫星位点LOH与P57^kip2mRNA和蛋白表达均无关联性。5．肝癌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原发性HCC的发生中起重要作用。2．与P57^kip2基因同一区域的3个微卫星位点未出现LOH，提示P57^kip2表达异常与该3个微卫星位点无关，可能存在其它机制。更多还原

【Abstract】 Objective: To study the loss of hererozygosity（LOH） of P57^kip2 and expressions of P57^kip2 mRNA & protein in human hepatocellular carcinoma （HCC） in order to explore the molecular mechanism of hepatocarcinogenesis.Methods: LOH of three microsatellite loci（D11S1397 D11S1318 and D11S4046） were detected where the P57^kip2 gene 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 detected: LOH was not identified in 30 cases on three microsatellite locies. 2. Expression of P57^kip2 mRNA: There were no expressions in normal liver tissue. Expressions both in pericancerous cirrhosis and hepatocellular carcinoma were 26. 67%（8/30）. There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 3. Expression of P57^kip2 protein: There were no expressions in normal liver tissue. Expressions in precancerous cirrhosis and hepatocellular carcinoma were 56. 67% （17/30） and 63.33 %（ 19/30） respectively. There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 4. There was no correlation between the LOH of three locies and the expression of P57^kip2mRNA; there was no correlation between the LOH of three locies and the expression of P57^kip2 protein. 5. There was positive correlation between expression of P57^kip2 mRNA and expression of P57^kip2 protein.Conclusion: 1. The disorder expressions of P57^kip2 mRNA and protein indicated that P57^kip2 may 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.更多还原