携带凋亡素的重组腺病毒激活AMPK诱导人膀胱癌EJ-1细胞S期阻滞  

AMPK activated by recombinant adenovirus carrying Apoptin induces S-phase arrest of human bladder cancer EJ-1 cells

作　　者：芦静 尚超 张翠玲 葛晨晨 李霄 金鑫[1] 金宁一 LU Jing;SHANG Chao;ZHANG Cui-ling;GE Chen-chen;LI Xiao;Jin Xin;JIN Ning-yi(Agricultural College,Yanbian University,Yanji,Jilin 133002,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130122,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]中国农业科学院长春兽医研究所,长春130122

出　　处：《军事医学》2021年第11期828-833,共6页Military Medical Sciences

基　　金：“科技助力经济2020”重点专项(SQ2020YFF0417940);吉林省青年科技人才托举工程项目(QT202111)。

摘　　要：目的探讨携带凋亡素(Apoptin)的重组腺病毒Ad-VT对人膀胱癌EJ-1细胞的杀伤作用及其细胞周期调节机制。方法取对数生长期的EJ-1细胞分别给予不同感染剂量的腺病毒载体对照(Ad-T)或Ad-VT处理,通过CCK-8法和克隆形成实验检测Ad-T和Ad-VT对EJ-1细胞增殖活力的影响;流式细胞术检测细胞周期分布;Western印迹检测S期调控蛋白CDK2、CyclinA2和p21,以及通路蛋白腺苷酸活化蛋白激酶(AMPK)和p-AMPK的表达水平。结果Ad-T和Ad-VT显著降低EJ-1细胞存活率,诱导EJ-1细胞S期阻滞,伴p21蛋白表达增加、CyclinA2和CDK2蛋白表达降低等S期检查点改变,且Ad-VT作用强于Ad-T。Ad-T和Ad-VT显著上调p-AMPK蛋白表达,AMPK抑制剂Dorsomorphin逆转了Ad-T和Ad-VT所致EJ-1细胞S期阻滞,而过表达AMPK处理促进了上述变化。结论Ad-VT具有抗膀胱癌作用,通过腺病毒载体和Apoptin两方面作用激活AMPK蛋白诱导人膀胱癌EJ-1细胞S期阻滞。Objective To investigate the anti-tumor effects of a recombinant adenovirus expressing Apoptin,designated as Ad-human telomerase reverse transcriptase(hTERT)-E1 a-Apoptin(Ad-VT),against human bladder cancer EJ-1 cells and the mechanisms of cell cycle regulation.Methods EJ-1 cells in the logarithmic growth phase were treated with different doses of Ad-VT or a control adenovirus Ad-T.The effect of Ad-T or Ad-VT on the proliferation of EJ-1 cells was detected with the CCK-8 method and clone formation assay.The distribution of cell cycles was detected by flow cytometry.The expressions of proteins that regulated S phase as well as AMPK/p-AMPK expression were detected by Western blotting.Results Both Ad-T and Ad-VT significantly decreased the proliferation rate of EJ-1 cells,and induced S-phase arrest,which was accompanied by corresponding alterations in S phase regulatory proteins,including up-regulation of p21 expression and down-regulation of CyclinA2 and CDK2 expressions.Ad-T or Ad-VT significantly up-regulated the expression of p-AMPK while the AMPK inhibitor Dorsomorphin reversed the S phase arrest of EJ-1 cell cycle.Overexpression of AMPK promoted the above changes caused by Ad-T and Ad-VT.Conclusion Ad-VT has anti-tumor effects on bladder cancer cells from both the adenovirus vector and Apoptin by inducing S phase arrest through AMPK activation.

关 键 词：膀胱癌 凋亡素 腺病毒 细胞增殖 细胞周期 

分 类 号：R737.14[医药卫生—肿瘤]