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甲基化紊乱与胃癌的发生有关,近年来甲基化已成为肿瘤研究的热点。目的:探讨在DNA甲基转移酶抑制剂5-氮脱氧胞苷(5-aza-2’-deoxycytidine,5-aza-dC)的化学干预下,不同分化胃癌细胞系的抑癌基因、癌基因和与凋亡相关的基因与甲基化调控的关系,并辅以流式细胞仪检测细胞周期的变化。方法:培养高分化、中分化和未分化胃癌细胞系MKN-45、MKN-28和:HGC-27,分别以不同浓度的5-aza-dC干预细胞。提取细胞的RNA,用逆转录聚合酶链反应(RT-PCR)方法检测p16INK4A、p21WAF1、p73、c-myc、c-Ha-ras、survivin和死亡相关蛋白激酶(DAP-ki-nase)等多种基因的表达情况;同时以流式细胞仪分析细胞周期的变化。结果:抑癌基因中,MKN-45和HGC-27细胞中有p16INK4A表达,用5-aza-dC干预后其表达增强,MKN-45细胞的p16INK4A在5-aza-dC 10μmcl/L 24 h、2 μmol/L72 h和5μmol/L 72 h组表达增强,HGC-27细胞的p16INK4A在5 μmol/L 24 h和10 μmol/L 24 h组表达也有明显增强;p21WAF1、p73无明显变化。癌基因中c-myc、c-Ha-ras变化不明显。与凋亡相关的基因中,MKN-45细胞的survivin在5-aza-dC处理后表达增强,但HGC-27细胞的DAP-kinase无明显变化。结论:在不同分化的人胃癌细胞系中,甲基化修饰对抑癌基因、癌基因以及与凋亡相关基因的调控
Methylation disorder and the occurrence of gastric cancer, methylation in recent years has become a hot tumor research. OBJECTIVE: To investigate the effect of 5-aza-2-deoxycytidine (5-aza-dC) on tumor suppressor genes, oncogenes And the relationship between apoptosis-related genes and methylation regulation, supplemented by flow cytometry to detect cell cycle changes. Methods: The well-differentiated, moderately differentiated and undifferentiated gastric cancer cell lines MKN-45, MKN-28 and HGC-27 were cultured and the cells were treated with different concentrations of 5-aza-dC. Cell RNA was extracted and many other proteins such as p16INK4A, p21WAF1, p73, c-myc, c-Ha-ras, survivin and DAP-kinases were detected by reverse transcriptase polymerase chain reaction The gene expression was analyzed by flow cytometry. The cell cycle was analyzed by flow cytometry. Results: The expression of p16INK4A in MKN-45 and HGC-27 cells was enhanced by 5-aza-dC, while the expression of p16INK4A in MKN-45 cells was significantly increased after 5-aza-dC 10μmcl / The expression of p16INK4A in HGC-27 cells was also significantly increased in 24 h and 5 μmol / L 24 h and 10 μmol / L 24 h groups; p21WAF1 and p73 had no significant changes. Oncogene c-myc, c-Ha-ras change was not obvious. In apoptosis-related genes, survivin of MKN-45 cells increased after 5-aza-dC treatment, but there was no obvious change of DAP-kinase in HGC-27 cells. CONCLUSION: Methylation modification plays a role in the regulation of tumor suppressor genes, oncogenes and apoptosis-related genes in different human gastric cancer cell lines