肝细胞癌和慢性肝病组织中p27^(kip1)基因表达及甲基化分析  

Expression and Methylation Analysis of p27^(kip1) in Human Hepatocellular Carcinoma and Chronic Liver Diseases

作　　者：雷普平[1] 申丽娟[1] 章宗籍[1] 李晋云[2] 熊昌庆[3] 张华献[1] 

机构地区：[1]昆明医学院病理教研室,云南昆明650031 [2]昆明医学院第二附属医院病理科,云南昆明650101 [3]昆明医学院第二附属医院肝胆外科,云南昆明650101

出　　处：《昆明医学院学报》2007年第2期12-16,共5页Journal of Kunming Medical College

基　　金：云南省自然科学基金(2000C0058M;2005C0045M);云南省教育厅科学研究基金(0011010)

摘　　要：目的研究肝细胞癌和慢性肝病中细胞周期调控因子p27kip1的表达和甲基化的关系.方法应用免疫组织化学、原位分子杂交技术分别检测正常肝组织、肝硬化、癌周肝硬化和肝细胞癌共68例标本中P27蛋白和p27mRNA的表达.应用甲基化特异性PCR技术检测正常肝组织、肝硬化和肝细胞癌共44例标本中p27基因甲基化情况.结果P27蛋白阳性率在正常肝组织(66.7%,4/6)、肝硬化(60.0%,6/10)和癌周肝硬化(50.0%,12/24)各组间差异没有统计学意义(P>0.05),p27mRNA阳性率在正常肝组织(83.3%,5/6)、肝硬化(70.0%,7/10)和癌周肝硬化(75.0%,18/24)各组间差异也没有统计学意义(P>0.05),但肝细胞癌组P27蛋白(21.4%,6/28)和mRNA(25.0%,7/28)的阳性率与各组比较均显著下降(P<0.05).P27蛋白阳性信号在正常肝组织和肝硬化组主要定位于胞核,而癌周肝硬化组和肝细胞癌组主要定位于胞浆.正常组与肝硬化合并组P27蛋白和p27mRNA表达具有相关性,癌周肝硬化组和肝细胞癌组P27蛋白与p27mRNA表达不具有相关性.44例样本中检出1例肝细胞癌的p27基因发生甲基化,其蛋白和mRNA表达均呈阴性.结论P27蛋白及p27mRNA表达下降或缺失可能参与了肝细胞癌的发生和发展,p27基因甲基化可能导致p27mRNA的失转录.Objective To study the expressions and the methylation of p27^kip1 in human hepatoceUular carcinoma （HCC） and chronic liver diseases. Methods P27 protein and p27 mRNA were detected by immunohistochemical staining and in situ hybridization （ISH） respectively in 68 cases of normal liver, liver cirrhosis, pericancerous cirrhosis and hepatoceUular carcinoma. The hypermethylation of p27^kip1 was detected by methylation - specific PCR （MSP） in 44 cases. Results The positive rates of P27 protein were 66.7% （4/6） in normal liver, 60. 0% （6/10） in liver cirrhosis, 50.0% （12/24） in pericancerous cirrhosis and 21.4% （6/28） in HCC. There were no statistical significance among normal liver, liver cirrhosis and pericancerous cirrhosis （P 〉 0.05） , but it was significantly decreased in HCC than those in other groups （ P 〈 0.05）. The positive rates of p27^kip1 mRNA were 83.3% （5/6） in normal liver, 70.0% （7/10） in liver cirrhosis, 75.0% （18/24） in peri-cancerous cirrhosis and 25.0% （7/28） in HCC. As the same as P27 protein, there was no statistical significance among normal liver, liver cirrhosis and pericancerous cirrhosis （ P 〉 0. 05 ） , but it was also significantly decreased in HCC those in other groups （P 〈 0. 05）. In addition, the positive signals of P27 protein were mainly located in cytoplasm in normal liver and liver cirrhosis while it was major located in nuclear in pericancerous cirrhosis and HCC. There was significant correlation between the expression of P27 protein and p27 mRNA in the integrated group of normal liver and liver cirrhosis, however, no significant correlations in both pericancerous cirrhosis and HCC. By methylation - specific PCR （MSP） , one of the 44 cases showed hypermethylation-both P27 protein and p27 mRNA were negative in this HCC case. Conclusion The reduction or loss of P27 protein and p27 mRNA potentially involved in hepatocarcinogenesis. The hypermethylation of p27 might lead to the loss of p27 mRNA transcription.

关 键 词：肝细胞癌/肝肿瘤 p27/p27^kip1 免疫组织化学 原位分子杂交 甲基化特异性PCR/MSP 

分 类 号：R735.7[医药卫生—肿瘤]