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目的探讨使用5杂氮2′脱氧胞苷(5DC)诱导肝癌细胞株癌/睾丸抗原(cancer/testis,CT)基因的可行性。方法使用5DC处理前后,用RTPCR方法检测肝癌细胞株QGY7703、HepG2215、HepG2、SMMC7721、HLE、BEL7402、Huh7、BEL7404中CT抗原基因MAGEA1、A3、A6和A10表达;Southern杂交检测细胞株基因组DNA去甲基化状态。结果CT抗原家族中MAGEA1在QGY7703、SMMC7721、BEL7402、HLE、BEL7404中表达,MAGEA3在HepG2、HLE中表达,MAGEA4和MAGEA10表达均为阴性,而MAGEA6表达均为阳性;使用5DC后,MAGEA4在QGY7703、HLE中出现表达,MAGEA10在HepG2和HLE出现表达。去甲基化后基因组DNA相对去甲基化程度明显升高(t=-4.966,P<0.01)。结论尽管5DC使肝癌细胞株基因组去甲基化程度明显升高,但不能有效地诱导CT抗原基因广泛表达。
Objective To investigate the feasibility of using 5-azacytidine 2-deoxycytidine (5DC) to induce cancer / testis (CT) gene in hepatocellular carcinoma cell lines. Methods The expression of CT antigen genes MAGEA1, A3, A6 and A10 in QGY7703, HepG2215, HepG2, SMMC7721, HLE, BEL7402, Huh7 and BEL7404 were detected by RTPCR before and after treatment with 5DC. Southern blotting was used to detect the genomic DNA Base state. Results MAGEA1 was expressed in QGY7703, SMMC7721, BEL7402, HLE and BEL7404, MAGEA3 was expressed in HepG2 and HLE, but both of MAGEA4 and MAGEA10 were negative, while MAGEA6 was all positive. After 5DC, MAGEA4 was expressed in QGY7703, Expression in HLE, expression of MAGEA10 in HepG2 and HLE. Demethylation of genomic DNA relative demethylation was significantly increased (t = -4.966, P <0.01). Conclusion Although 5DC can significantly demethylate the hepatoma cell lines, it can not effectively induce the widespread expression of CT antigen genes.