Survivin介导的条件复制型溶瘤腺病毒表达IL-24蛋白对人肝癌细胞PLC作用机制探究

Antitumor Mechanism of Survivin Mediated Conditionally Replicative Oncolytic Adenoviruse Expressing IL-24 in Human Hepatocellular Carcinoma Cell Line PLC

条件复制型溶瘤腺病毒Ad.sp-E1A_((△24)-IL-24对人肝癌细胞株PLC的作用机制尚不清楚。采用流式细胞仪检测PLC经PBS、Ad.sp-E1A_((△24)以及Ad.sp-E1A_((△24)-IL-24处理48h后细胞周期的变化;利用Western blot检测PLC经PBS、Ad.sp-E1A_((△24)以及Ad.sp-E1A_((△24)IL-24处理24 h、48h后细胞中凋亡信号相关蛋白表达量的变化。实验结果表明:Ad.sp-E1A_((△24)和Ad.sp-E1A_((△24)-IL-24能将PLC细胞周期阻滞在S期,其值分别为43.74%±6.61%,49.48%±7.60%,两者与未感染病毒的对照组(18.91%±1.83%)进行统计学差异分析,P值均<0.05;同时,Western blot检测发现随着处理时间的增加,病毒处理组中CHOP的表达量、caspase 12的剪切水平及STAT3、p38 MAPK的磷酸化水平都有明显提高,尤其以Ad.sp-E1A_((△24)-IL-24处理组较为明显。SOCS3的表达量病毒处理组与对照组没有显著差异。上述结果表明重组腺病毒Ad.sp-E1A_((△24)-IL-24能有效阻滞PLC细胞周期在S期,并可能通过内质网压力、STAT3与MAPK信号通路等多种途径诱发PLC细胞凋亡。

Antitumor mechanism in human hepatocellular carcinoma cell line PLC induced by conditionally replicative oncolytic adenoviruse（Ad.sp-E1A_（（△24））-IL-24） is not clear.PLC was treated with PBS,Ad.sp-E1A_（（△24）） or Ad.sp-ElA_（（△24））-IL-24 for 48 h,respectively.The cell cycle was detected by flow cytometry（FCM）.Western blot detected the changes of apoptosis-related protein expression.FCM confirmed that both Ad.sp-ElA_（（△24）） and Ad.sp- E1A_（（△24））-IL-24 can block PLC cells in S phase,of which the percentages were 43.74%±6.61%and 49.48%±7.60% compared with 18.91%±1.83%of control（P0.05）.Western blot indicated both Ad.sp-E1A_（（△24）） and Ad.sp-E1A_（（△24））-IL -24 can increase the expression of CHOP,cleavaged caspase 12,p-STAT3 and p-p38 MAPK.Obviously,the virus Ad.sp-E1A_（（△24））-IL-24 had the stronger ability of inducing the above proteins,but both of the viruses had no effect on inducing expression of SOCS3.In conclusion,the recombinant oncolytic adenovirus blocked PLC cells in S phase effectively,and induced apoptosis of PLC cells directly related to endoplasmic reticulum stress,STAT3 and MAPK pathways.