摘要
目的探讨莱菔硫烷(SFN)对胃癌细胞SGC-7901 RASSF1A甲基化及表达有何影响。方法体外培养胃癌细胞SGC-7901细胞,用0,10μM,30μM,50μM的SFN与SCG-7901细胞孵育24～96h,采用甲基化特意性PCR检测RASSF1A基因启动子区域的甲基化改变情况;提取EGCG处理后SGC-7901细胞的RNA,用逆转录PCR检测RASSF1A的表达水平。结果静息状态下,SCG-7901细胞可检测出RASSF1A基因启动子区域有明显的甲基化条带,当与50μM SFN孵育96h后,甲基化水平有所减弱,而非甲基化基因增多。SFN也能以剂量依赖性与时间依赖性方式促进SGC-7901细胞RASSF1A基因mRNA的表达。结论SFN能使RASSF1A基因去甲基化,并上调非甲基化基因的表达水平,这可能是其抗肿瘤的重要机制。
Objective To investigate the effect of sulforaphane（SFN） on the methylation of RASSFIA and mRNA expression level in a cell line of SGC-7901. Methods The cultured SGC-7901 cells were incubated with SFN for 24 -96h. After that, DNA was extracted, and the methylation in RASSF1A promoter region was determined by methylation-specific PCR. The mRNA level of RASSF1A was detected by RT-PCR. Results PCR results showed non-methylation band and methylation band was detected when SGC-7901 cells was under quiescent condition. When the cells were incubated with 50μM SFN, the methylation band level was decreased, and the non-methylation was DNA increased.RT-PCR showed the mRNA level of RASSF1A increased in dose and time-dependent manner after SGC-7901 cells was treated with SFN. Conclusion SFN demethylation of RASSFIA and increase its mRNA may be one of the mechanisms of its antineoplastic activity.
出处
《中国现代医生》
2011年第7期8-9,20,共3页
China Modern Doctor
