shRNA干扰PARG对大肠癌CT26细胞粘附因子表达的影响  

Influence of shRNA interfering with PARG on the expression of cell adhesion fact

作　　者：颜佳欣[1] 王娅兰[1] 吴伟强[1] 潘娟[1] 

机构地区：[1]重庆医科大学基础医学院病理学教研室分子医学与肿瘤研究中心,重庆400016

出　　处：《重庆医科大学学报》2011年第6期641-645,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:NSFC30870946)

摘　　要：目的:探讨小鼠大肠癌CT26株中聚(腺苷二磷酸核糖)水解酶[Poly(ADP-ribose)glycohydrolase,PARG]对细胞间粘附因子表达的影响及其可能的机制。方法:以慢病毒为载体对CT26细胞株中PARG基因进行RNA沉默干扰,并筛选出稳定表达株。Realtime-PCR法检测沉默效果。Western blot法检测CT26细胞PARG、PARP、NF-κB和Pi-AKT(473)I、CAM-1、P-selectin的蛋白表达。结果:实验组与对照组(包括阴性对照组、空载体对照组)相比,PARG、PARP、Pi-AKT(473)、NF-κBI、CAM-1、P-selectin表达经统计学分析差异均有统计学意义(P<0.05)。结论:在大肠癌细胞株中,抑制PARG可以通过调节与NF-κB有关通路,进一步调节ICAM-1、P-selectin等细胞粘附因子。提示PARG在肿瘤癌栓形成过程中可能具有重要作用。Objective：To clarify the possible mechanism of how PARG and PARP are to regulate intercellular adhesion factor through relevant signal pathways.Methods：Lentivirus carried shRNA interfered with the expression of PARG and steadily silenced cell lines.Realtime-PCR was used for testing the strength of shRNAi and Western blot was used to test the expressions of PARG,PARP,NF-κB,and Pi-AKT（473）,and LY249002 was served as Pi-AKT inhibitor.Results：The expressions of PARG,PARP,and NF-κB,ICAM-1,P-selectin in shRNAi group were weaker than those in the control groups（blank control and transfected with empty vector control group）;whereas Pi-AKT increased in the shRNAi group（P〈0.05）.Conclusion：The results suggest that PARG gene silencing may regulate NF-κB related signaling pathway and the intercellular adhesion factors such as ICAM-1 and P-selectin,which are reduced.

关 键 词：PARG 慢病毒转染 AKT磷酸化 细胞粘附 

分 类 号：R364[医药卫生—病理学]