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目的:研究IMP3在原发性肝细胞肝癌中的表达意义以及基因甲基化同IMP3表达的关系。方法:免疫组化检测162例肝细胞肝癌组织中IMP3的表达。分析临床因素与IMP3表达水平的相关性,并随访所选162例术后患者生存情况作生存分析。提取正常肝组织、肝癌组织的DNA,通过甲基化特异性定量PCR测定IMP3启动子CpG岛甲基化状态。培养正常肝脏细胞L02和Huh-7肝癌细胞,通过Western blot检测IMP3蛋白的表达。提取两组细胞DNA,分别对IMP3基因启动子区域CpG岛进行甲基化状态检测。结果:原发性肝细胞肝癌组织中IMP3阳性率为67.90%;IMP3表达与患者年龄、肿瘤大小、分级、分期、AFP、早期复发均存在相关性;而与性别、HBsAg无明显相关性。随访显示IMP3阳性患者长期生存明显低于IMP3阴性者。CpG岛甲基化检测显示肝癌组织中P1和P2位点高度去甲基化,而正常肝脏组织中P1和P2则处于甲基化状态。正常肝脏细胞IMP3表达阴性,Huh-7肝癌细胞IMP3表达强阳性。正常肝细胞P1和P2位点处于高度甲基化状态,而Huh-7肝癌细胞P1和P2位点处于去甲基化状态。结论:IMP3在原发性肝细胞肝癌中具有高表达,IMP3高表达提示不良预后;基因的甲基化同IMP3的表达存在一定的相关性。
Objective: To study the expression of IMP3 in primary hepatocellular carcinoma and its relationship with the expression of IMP3. Methods: The expression of IMP3 in 162 hepatocellular carcinoma tissues was detected by immunohistochemistry. The correlation between clinical factors and IMP3 expression level was analyzed. Survival analysis of the survival of 162 selected patients was followed up. The DNA of normal liver tissues and liver cancer tissues was extracted, and the methylation status of CpG island of IMP3 promoter was determined by methylation-specific quantitative PCR. Normal liver cells L02 and Huh-7 hepatoma cells were cultured and the expression of IMP3 protein was detected by Western blot. Two sets of cell DNA were extracted and the methylation status of CpG island in promoter region of IMP3 gene was detected. Results: The positive rate of IMP3 in primary hepatocellular carcinoma was 67.90%. The expression of IMP3 correlated with age, tumor size, stage, AFP and early recurrence, but not with sex and HBsAg. Follow-up showed that the long-term survival of IMP3-positive patients was significantly lower than that of IMP3-negative patients. The methylation of CpG island showed that the P1 and P2 sites in HCC tissues were highly demethylated while P1 and P2 in normal liver tissues were methylated. The expression of IMP3 in normal liver cells was negative, and the expression of IMP3 in Huh-7 hepatoma cells was strongly positive. The P1 and P2 sites of normal hepatocytes are highly methylated while the P1 and P2 sites of Huh-7 hepatoma cells are in a demethylated state. Conclusion: IMP3 is highly expressed in primary hepatocellular carcinoma and high expression of IMP3 is associated with poor prognosis. There is a correlation between methylation and IMP3 expression.