Ku70乙酰化修饰介导曲古霉素A(TSA)促结肠癌细胞凋亡的作用机制研究  

The mechanism of Ku70 acetylation modification mediating apoptosis of colon cancer cells induced by trichostatin A( TSA)

作　　者：孟瑾[1,2] 刘新利 车玲 陈明 吴漫[1] 马晨珂 赵冠人 Meng Jin;Liu Xinli;Che Ling;Chen Ming;Wu Man;Ma Chenke;Zhao Guanren(Department of Pharmacy,the Eighth Medical Center of Chinese PLA General Hospital,Beijing 100091,China;Department of Oncology,Tangdu Hospital,Air Force Medical University,Shaanxi Xi'an 710038,China;College of Life,Northwestern Polytechnical University,Shaanxi Xi'an 710072,China)

机构地区：[1]解放军总医院第八医学中心药剂科,北京100091 [2]空军军医大学唐都医院肿瘤科,陕西西安710038 [3]西北工业大学生命学院,陕西西安710072

出　　处：《现代肿瘤医学》2020年第3期355-360,共6页Journal of Modern Oncology

基　　金：陕西省自然科学基础研究计划项目(编号:2017JM8034);解放军总医院第八医学中心课题基金项目(编号:2016MS-016)

摘　　要：目的:探讨Ku70乙酰化修饰介导曲古霉素A(TSA)促结肠癌细胞凋亡的作用和机制。方法:选取结肠癌HCT116细胞和SW620细胞,体外培养并采用浓度梯度TSA处理。MTT法观察TSA对细胞的IC 50和活力的影响;Western blot和免疫荧光染色观察TSA对结肠癌HCT116细胞和SW620细胞中Ku70、acetyl-Ku70蛋白水平及胞内分布的影响;流式细胞术检测TSA对结肠癌HCT116细胞和SW620细胞凋亡的影响;Western blot检测TSA对结肠癌HCT116细胞和SW620细胞中凋亡相关蛋白Caspase-3、Bax和Bcl-2表达的影响。结果:MTT结果显示TSA可浓度依赖性抑制结肠癌HCT116细胞和SW620细胞活力且其IC 50值分别为1.023μmol/L和1.076μmol/L(P<0.05);Western blot和免疫荧光染色结果显示TSA可显著上调结肠癌HCT116细胞和SW620细胞中acetyl-Ku70的蛋白水平并促进其核转入(P<0.05);流式细胞术结果表明TSA可促进结肠癌HCT116细胞和SW620细胞凋亡(P<0.05);此外,Western blot结果显示TSA可明显上调结肠癌HCT116细胞和SW620细胞中凋亡蛋白Caspase-3和Bax的表达,下调抗凋亡蛋白Bcl-2的表达(P<0.05)。结论:结肠癌HCT116细胞和SW620细胞中,TSA可能通过增强Ku70乙酰化并促进其核转入,发挥促凋亡作用,为以Ku70翻译后修饰调控为靶点的肿瘤治疗提供新策略和理论依据。Objective:To investigate the effect and mechanism of Ku70 acetylation modification on the apoptosis of colon cancer cells induced by trichostatin A(TSA).Methods:Colon cancer HCT116 cells and SW620 cells were selected and cultured in vitro and treated with concentration gradient TSA.MTT assay was used to detect the IC 50 and cell viability treated with TSA.Western blot and immunofluorescence staining assays were used to detect the effects of TSA on the protein level and intracellular distribution of Ku70 and acetyl-Ku70.The flow cytometry detected the effect of TSA on apoptosis of colon cancer HCT116 cells and SW620 cells.The effects of TSA on the expression of apoptosis-related proteins,including Caspase-3,Bax and Bcl-2 in colon cancer HCT116 cells and SW620 cells were detected by Western blot.Results:MTT assay results showed that TSA could inhibit the activity and growth of colon cancer HCT116 and SW620 cells in a concentration-dependent manner with an IC 50 value of 1.023μmol/L and 1.076μmol/L(P<0.05).Western blot and immunofluorescence staining results showed that TSA could significantly up-regulate the protein level of acetyl-Ku70 in colon cancer HCT116 and SW620 cells and promote its nuclear transduction(P<0.05).Flow cytometry results showed that TSA could significantly promote colon cancer HCT116 and SW620 cells apoptosis(P<0.05).In addition,Western blot results showed that TSA could significantly up-regulate the apoptosis protein Caspase-3 and Bax in colon cancer HCT116 and SW620 cells,and down-regulate the expression of anti-apoptosis protein Bcl-2(P<0.05).Conclusion:In the colon cancer HCT116 and SW620 cells,TSA may play a role in targeting the Ku70 post-translational modification regulation by mediating the acetylation of the proto-oncogene Ku70 and promoting its nuclear transduction to promote its apoptosis,and providing a new strategy and theoretical basis for the anti-treatment of colon cancer cells.

关 键 词：结肠癌 曲古霉素A(TSA) 乙酰化 凋亡 

分 类 号：R735.35[医药卫生—肿瘤]