莱菔硫烷对鼻咽癌RASSF1A基因甲基化及表达的影响  

Effect of sulforaphane on RASSF1A methylation and gene expression in nasopharyngeal carcinoma cells

作　　者：余盛富[1] 李谨[1] 张功秀[1] 

机构地区：[1]十堰市人民医院耳鼻喉科,湖北十堰442000

出　　处：《中国热带医学》2011年第8期935-937,共3页China Tropical Medicine

摘　　要：目的探讨莱菔硫烷(SFN)对鼻咽癌CNE-2细胞RASSF1A甲基化及蛋白表达水平的影响。方法体外培养鼻咽癌CNE-2细胞,用0,5、10、30μM的SFN与其孵育24～96h,用MTT法检测细胞的增值情况;流式细胞术分析细胞周期的改变;采用甲基化特异性PCR检测RASSF1A基因启动子区域的甲基化改变情况;提取SFN处理后细胞的RNA,用逆转录PCR检测RASSF1A的表达水平,并用Western blot检测处理前后RASSF1A蛋白的表达。结果 SFN能以时间和剂量依赖性方式抑制CNE-2细胞增值,并能使细胞周期阻滞于S期和G2/M期。随着SNF浓度的增加,甲基化RASSF1A水平逐渐减弱,非甲基化RASSF1A水平逐渐增多。同时SFN能明显上调RASSF1A的mRNA和蛋白表达水平。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,从而抑制细胞的增值和分化而具有抗肿瘤的效应。Objective To investigate the effect of sulforaphane（SFN） on RASSF1A methylation,mRNA and protein expression in a nasopharyngeal carcinoma cell line of CNE-2.Methods The cultured CNE-2 cells were incubated with 0,5μM,10μM,30μM of SFN for 24～96h.After that,MTT was used to assess cell proliferation of CNE-2.Cell cycle redistribution was detected by flow cytometry.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,and the protein level of RASSF1A was detected by Western blot.Results SFN could inhibit CNE-2 cells proliferation in time in a dose-dependent manner.SNF also arrested CNE-2 cellls in G2/M phase of cell cycle.PCR results showed non-methylation band of RASSF1A became weaker after incubated by increased concerntration of SFN,and the methylation band became stronger at mRNA and protein level.Conclusion SFN demethylation of RASSF1A and increase its mRNA may be one of the mechanisms of its antineoplastic activity.

关 键 词：莱菔硫烷 胃癌 Ras相关区域家族1基因 

分 类 号：R739.61[医药卫生—肿瘤]