5-氮杂-2-脱氧胞苷及曲古抑菌素A对人胃癌SGC-7901细胞生长及CHFR基因表达水平的影响  

The effect of combination 5-Aza-2'-deoxycytidine with or without trichostatin A on the growth of cancer cells and CHFR expression in gastric cancer cell line SGC-7901

作　　者：刘林青[1,2] 胡世莲[1,2] 沈干 徐维平 沈建军[3] 姜晓东[4] 黄大兵[4] 黄毕林[1,2] 方中良[1,2] 王海[1,2] 唐杨琛[1,2] 

机构地区：[1]安徽医科大学附属省立医院老年病科,安徽合肥230001 [2]安徽省老年病研究所,安徽合肥230001 [3]安徽医科大学附属省立医院肿瘤放疗科,安徽合肥230001 [4]安徽医科大学附属省立医院肿瘤化疗科,安徽合肥230001

出　　处：《中国癌症杂志》2012年第8期573-577,588,共6页China Oncology

基　　金：国家自然科学基金项目资助(No:81071808);安徽省自然科学基金(No:1208085QH169);安徽省卫生厅资助项目(No:09A083);安徽省卫生厅资助项目(No:10021403080)

摘　　要：背景与目的:抑癌基因甲基化具有可逆性,相关药物可逆转甲基化使失活的基因重新表达。本研究观察去甲基化制剂5-氮杂-2-脱氧胞苷(5-Aza-dC)和组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)对体外培养的人胃癌SGC-7901细胞活性及CHFR基因mRNA和蛋白表达水平的影响。方法:使用不同浓度的5-Aza-dC和(或)TSA处理SGC-7901细胞,分为对照组、TSA组(250 nmol/L)、5-Aza-dC组(5μmol/L)及TSA(250 nmol/L)联合5-Aza-dC(5μmol/L)组等4个组,采用台盼蓝染色法检测细胞活性,并利用RT-PCR及蛋白质印迹法(Westernblot)方法检测CHFR基因mRNA和蛋白表达的水平。结果:5-Aza-dC和TSA作用24、48、60和72 h后SGC-7901细胞活性均被抑制,且抑制率呈浓度和时间依赖性,两药联合作用更明显;SGC-7901细胞经5-Aza-dC单独或与TSA联合干预后,能提高CHFR基因mRNA和蛋白表达水平,差异有统计学意义(P<0.05)。结论:5-Aza-dC和(或)TSA均能抑制细胞活性,提高CHFR基因mRNA和蛋白表达水平,两药联合的效果比单药明显增加,为临床胃癌患者的化疗方案提供了实验依据。Background and purpose： Methylation of anti-oncogene has reversibility by the related drugs for reversal of methylation inactivation of reprogramming of gene expression. Demethylating agent 5-Aza-2＇-deoxycytidine and histone deacetylase inhibitors trichostatin A can reverse the methylation of gene re-expression. This study aimed to research on the effects of 5-Aza-2＇-deoxycytidine （5-Aza-dC） with or without trichostatin A （TSA） on the expression of CHFR gene and the growth of human gastric cancer cell line SGC-7901. Methods： SGC-7901 cells were treated with different 5-Aza-dC or （and） TSA concentrations, experimental were divided into the control group, the group with TSA （250 nmol/L）, the group with 5-Aza-dC （5μmol/L）, the group with 5-Aza-dC （5 μmol/L） and TSA （250 nmol/ L） in combination, Cell viability rates were detected by Trypan Blue, expression of CHFR was detected by the mRNA and protein expression levels of CHFR was detected by transcription polymerase chain reaction （RT-PCR） and Western blot. Results： 5-Aza-dC and （or） TSA both can reduce the growth of SGC-7901 cells after being treated with 24, 48, 60 and 72 h. The cell survival rate in the union medication group was significantly higher than that in the independent medication group. After treatment with 5-Aza-dC and （or） TSA, the hypermethylation of CHFR gene in SGC-7901 ceils was reversed to an unmethylated pattern. Conelnsion： 5-Aza-dC and （or） TSA can inhibit the cell viability of SGC- 7901 ceils, reverse CHFR gene methylation levels, recover its mRNA and protein expression. In the union medication group, it was much more effective than that in the independent medication group to reduce methylation level. It provides experimental basis for trichostatin A and （or） 5-Aza-2＇-deoxycytidine in treating gastric cancer.

关 键 词：5-氮杂2-脱氧胞苷 曲古抑菌素A 胃癌 SGC-7901细胞 CHFR基因 

分 类 号：R735.2[医药卫生—肿瘤]