曲古抑菌素A抑制胃癌细胞增殖的分子机制  

Trichostatin A inhibits the proliferation of gastric cancer cells through the promotion of autophagy via p-Akt/p-mTOR pathway

作　　者：汤志明 谷春明 刘洋[1] 覃韦宁 李佳莉 杨萍[1] 杨方红 蔡云丽 武福云[1] 李珊[1] TANG Zhi-ming;GU Chun-ming;LIU Yang;QIN Wei-ning;LI Jia-li;YANG Ping;YANG Fang-hong;CAI Yun-li;WU Fu-yun;LI Shan(Institute of Basic Medical Science,Hubei University of Medicine,Shiyan 422000,Hubei,China)

机构地区：[1]湖北医药学院基础医学研究所

出　　处：《医学研究生学报》2020年第1期7-11,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金（81502637）;湖北省自然科学基金（2018CFB467）

摘　　要：目的曲古抑菌素A(TSA)是氧肟酸盐类组蛋白去乙酰化酶抑制剂,具有一定的抗肿瘤活性。文中主要探讨胃癌细胞中TSA通过p-AKT/p-mTOR途径抑制细胞增殖的分子机制。方法分别用不同浓度的TSA处理胃癌细胞SGC-7901,通过MTT实验检测TSA对细胞的抑制率。将细胞分为对照组(不做任何处理)、TSA处理组(200 ng/mL TSA)、p-AKT回补组(200 ng/mL TSA+8μg/mL SC79)和自噬抑制组(5 mmol/mL 3-甲基腺嘌呤+200 ng/mL TSA)。通过细胞免疫荧光染色检测对照组和TSA处理组内Lc3的蛋白表达分布。Western blot检测对照组、TSA处理组和p-AKT回补组中p-AKT、p-mTOR以及自噬相关蛋白Lc3、P62的相对表达水平。EdU和细胞计数实验检测对照组、TSA组、p-AKT回补组和自噬抑制组中细胞的增殖能力。结果处理24 h,TSA处理组Lc3在细胞质内形成大量颗粒状聚集物,荧光分布从初始的弥散状转变为致密状。EdU实验显示,与对照组相比,TSA处理组的绿色荧光显著减少。对照组、TSA处理组、自噬抑制组p-AKT的表达量分别为1.78±0.19、0.92±0.03和1.71±0.19,Lc3分别为0.21±0.01、0.79±0.06和0.55±0.10,与对照组相比,TSA处理组中p-AKT相对表达水平均降低,Lc3表达升高(P<0.05)。3组p-mTOR分别为0.80±0.16、0.45±0.04和0.98±0.16,与对照组相比,TSA处理组中p-mTOR相对表达水平均降低(P<0.05)。3组P62分别为1.17±0.15、0.48±0.08和0.77±0.10,与对照组相比,TSA处理组中P62相对表达水平均降低(P<0.05)。与TSA处理组比较,p-AKT回补组p-AKT、p-mTOR、P62表达升高(P<0.05),Lc3表达降低(P<0.05)。与对照组相比,TSA处理组的细胞生长曲线抑制效应最明显,而p-AKT回补组和自噬抑制组较TSA处理组细胞生长曲线有部分回升。结论TSA可以通过抑制p-AKT/p-mTOR途径,促进自噬,进而抑制胃癌细胞的增殖。Objective Trihostatin A(TSA)is a histone deacetylase inhibitor of oxime salts,which has certain anti-tumor activity.This article mainly investigates the molecular mechanism of TSA inhibiting cell proliferation through p-AKT/p-mTOR pathway in gastric cancer cells.Methods Gastric cancer cell line-SGC-7901 were treated with TSA of different concentrations,and the inhibition rate of TSA on the cells was detected by MTT assay.The cells were divided into control group(without any treatment),TSA-treated group(200ng/ml TSA),p-AKT covering group(200 ng/mL TSA+8μg/mL SC79)and autophagy inhibition group(5 mmol/mL 3-methyladenine+200 ng/mL TSA).The protein expression distribution of Lc3 in control and TSA group were detected by cell immunofluorescence staining.The relative expression levels of p-AKT,p-mTOR and autophagy related proteins Lc3 and P62 in control group,TSA group and p-AKT covering group were detected by Western blot.The proliferation of cells in control group,TSA group,p-AKT covering group and autophagy inhibition group were measured by EdU and cell count assay.Results After 24h of treatment,Lc3 in TSA group formed a large number of granular aggregates in the cytoplasm,and the fluorescence distribution changed from the initial diffuse to dense.The TSA group showed a significant reduction in green fluorescence compared with the control group in the EdU experiment.The expression levels of p-AKT in the control group,TSA group and the autophagy inhibition group were 1.78±0.19,0.92±0.03 and 1.71±0.19,respectively,and Lc3 were 0.21±0.01,0.79±0.06 and 0.55±0.10,respectively.Compared with the control group,the relative expression level of p-AKT in the TSA group all decreased,while the expression level of Lc3 increased(P<0.05).p-mTOR in the three groups was 0.80±0.16,0.45±0.04 and 0.98±0.16,respectively.Compared with the control group,the relative expression level of p-mTOR in the TSA group all decreased(P<0.05).P62 in the three groups was 1.17±0.15,0.48±0.08 and 0.77±0.10,respectively.Compared with the cont

关 键 词：曲古抑菌素 蛋白激酶B 自噬 增殖 

分 类 号：R36[医药卫生—病理学]