5-氮杂-2-脱氧胞苷联合曲古抑菌素A对胃癌细胞SGC-7901MGMT表达及NF-κB活性的影响  

Effects of 5-Aza-dC combined with TSA on MGMT expression and NF-κB activity in gastric cancer cell line SGC-7901

作　　者：孟兰 程民 沈国栋 徐婷娟 刘林青[1,2] 唐杨琛 韩晓青 沈干 胡世莲 

机构地区：[1]安徽医科大学附属省立医院老年医学科,安徽省老年医学研究所 [2]肿瘤免疫与营养治疗安徽省重点实验室 [3]安徽省循证医学中心,安徽合肥230001

出　　处：《中国药理学通报》2014年第2期244-250,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 8107180);安徽省国际科技合作计划项目(No 1303063017);安徽省科技计划项目(No 10021403080)

摘　　要：目的研究5-氮杂-2-脱氧胞苷(5-Aza-2'-deoxycitydine,5-Aza-dC)及曲古抑菌素A(trichostatin A,TSA)对人胃癌细胞系SGC-7901细胞增殖、凋亡及移植瘤生长的影响,从体内、体外观察抑癌基因O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)表达,核转录因子NF-κB活性的改变,探讨5-Aza-dC联合TSA用于胃癌临床治疗的可行性。方法实验分对照组、5-Aza-dC组、TSA组和5-Aza-dC联合TSA组,利用CCK-8增殖试剂盒检测细胞增殖情况,流式细胞术分析细胞凋亡,RT-PCR检测各组MGMT mRNA表达,Western blot检测MGMT、NF-κB p65蛋白表达及NF-κB p65的磷酸化水平。结果 5-Aza-dC、TSA单独处理均可抑制SGC-7901细胞的生长和促进其凋亡,联合用药后效果更明显(P<0.05);与对照组相比,单药组移植瘤变化没有统计学意义(P>0.05),而联合用药组较单药组移植瘤变小;5-Aza-dC、TSA单独或联合处理SGC-7901细胞和裸鼠移植瘤后,与对照组比较,MGMT基因mRNA及蛋白表达水平均上调,联合用药比单独用药升高明显,NF-κB p65在各处理组中的表达量未发生变化,但其磷酸化程度较对照组下降,联合用药较单药下降更明显(P<0.05)。结论 5-Aza-dC与TSA发挥抑制胃癌生长的作用可能是通过抑制NF-κB的活性,提高MGMT基因的转录和表达而实现的,两药联合使用的抗癌效果更加明显。Aim To explore the potential effects of 5-Aza-2＇-deoxycitydine （5-Aza-dC） and trichostatin A （TSA） on the treatment of patients with gastric canc-er. Methods The inhibitory effects of 5-Aza-dC and TSA on SGC-7901 cell proliferation were assayed by u-sing CCK-8 kit. The apoptosis of SGC-7901 was deter-mined by Annexin V-FITC / PI staining method using flow cytometry. MGMT mRNA in vivo and vitro was de-termined by RT-PCR, and the MGMT, NF-KB p65 and p-NF-KB p65 protein expression were detected by Western blotting. Results The cell growth of human gastric carcinoma cell line SGC-7901 was significantly inhibited by 5-Aza-dC or TSA, and the combination of 5-Aza-dC and TSA resulted in a better inhibition.Compared with the control group, 5-Aza-dC and TSA group induced the apoptosis of SGC-7901 cells, with the apoptotic rate significantly higher in the combina-tion group than in single drug treatment group （P 〈 0.05 ）. There were significant increases in the expres-sion levels of MGMT mRNA and protein in 5-Aza-dC or TSA group, and the effects were consequently more pronounced in the combined group in vivo and vitro. Exposure of SGC-7901 cell line to 5-Aza-dC or TSA a-lone resulted in the down-regulation of p-NF-KB p65, and the effect of combined 5-Aza-dC with TSA was more remarkable than that of single drug. However, there was no significant difference in the protein ex- pression of NF - KB p 6 5 between those groups. Conelu-sion Exposure to 5-Aza-dC or TSA is shown to inhibit cell proliferation and induce apoptosis in human gastric carcinoma SGC-7901 cell line, which may be involved in the up-regulation of MGMT expression as well as the decreasing of NF-KB activity in vitro and vivo. Moreo-ver, 5-Aza-dC combined with TSA is more efficient than either 5-Aza-dC or TSA alone.

关 键 词：胃癌 细胞系 SGC-7901 5-氮杂-2-脱氧胞苷 曲古抑菌素A MGMT基因 核转录因子κB 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]