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目的:研究肝癌细胞株中DNA甲基化与HLAI类分子异常表达的相关性。方法:应用MSP技术对相关细胞系的HLA I类分子重链A、B、C位点启动子区域CpG岛的甲基化状态进行分析,Real-time PCR检测HLAI类分子重链mRNA水平的表达情况,Western blot检测RNA干扰后HLAI类分子重链表达情况。结果:在8株肝癌细胞系中HLA I类分子重链A、C位点启动子区域CpG岛存在甲基化;将启动子区域的DNA甲基化与相关基因表达数据相比较显示二者没有关联性;在RNA干扰DNA甲基化转移酶3a或3b的肝癌细胞系SMMC7721中,比较基因干扰前后HLA I类分子重链蛋白表达无显著变化。结论:在研究的肝癌细胞系中DNA甲基化没有参与调控肝癌中HLA I类分子的异常表达。
Objective: To investigate the relationship between DNA methylation and abnormal expression of HLA class I molecules in hepatocellular carcinoma cell lines. Methods: The methylation status of CpG islands in the promoter region of HLA class I heavy chain A, B, C sites in related cell lines was analyzed by MSP technique. The expression of HLAI heavy chain mRNA was detected by Real-time PCR Western blot was used to detect the expression of heavy chain of HLA class I molecules after RNA interference. Results: The methylation of CpG islands in the promoter region of HLA class I heavy chain in the eight HCC cell lines was found. Compared with the DNA methylation in the promoter region and the related gene expression data, There was no significant change in the expression of HLA class I heavy chain protein before and after interference by siRNA in SMMC7721 hepatocellular carcinoma cell line transfected with DNA methylated transferase 3a or 3b. CONCLUSIONS: DNA methylation is not involved in the regulation of HLA class I molecule abnormalities in hepatocellular carcinoma in the studied hepatocellular carcinoma cell lines.