RNA干扰LKB1基因对前列腺癌细胞增殖的影响  

The effects of LKB1 gene down- regulation induced by RNA in prostate carcinoma cell

作　　者：刘超颖[1] 唐伟[1] 蒋立[1] 熊晶[1] 邓玮[2] 

机构地区：[1]重庆医科大学附属第一医院,重庆400016 [2]重庆市第五人民医院,重庆400061

出　　处：《临床医药实践》2016年第7期531-535,共5页Proceeding of Clinical Medicine

摘　　要：目的:探讨LKB1/STKll(Liver kinase B1/Serine-Threonine Kinase II)基因沉默对前列腺癌细胞株PC-3中增殖相关因子细胞周期素D1(cyclin D1)、微小染色体维持缺陷蛋白2(Mcm2)和增殖细胞核抗原(PCNA)表达的影响及其可能机制。方法:将重组表达质粒LKB1-shRNA1(LKB1-shRNA1组)和阴性对照质粒sh NC(阴性对照组)采用脂质体介导法转染至前列腺癌细胞株PC-3中,并设未转染组,经G418筛选出稳定转染细胞,RT-PCR法检测转染前后细胞中LKB1,cyclin D1,Mcm2和PCNA基因mRNA的转录水平;Western blot法检测转染前后细胞中LKB1,cyclin D1,Mcm2和PCNA蛋白表达水平;CCK-8法检测转染后细胞的增殖能力。结果:与未转染组和阴性对照组相比,LKB1-shRNA1组细胞中LKB1,cyclin D1,Mcm2和PCNA的mRNA和蛋白表达水平均明显上调,LKB1-shRNA1组PC-3细胞的生长速度明显增加。结论:LKB1基因沉默上调前列腺癌细胞中cyclin D1,Mcm2和PCNA的表达,LKB1可作为研究前列腺癌细胞增殖分子机制的新靶点。Objective： To investigate the effect of Liver kinase B1 / Serine- Threonine Kinase II（ LKBl / STKll） gene silencing on expression of cyclin D1,Mcm2 and PCNA of human prostate carcinoma PC- 3 cells as well as the relevant mechanism. Methods： The shRNA expression plasmid of LKB1（ LKB1- shRNA1） was constructed and transfected to PC- 3 cellsby liposome- mediated transfection,while using the cells untransfected as blank group and those transfected with Vector as negativegroup,and the well- transfected cells were screened out using G418. The transcription levels of LKB1,cyclin D1,Mcm2 and PCNA mRNAs in the transfected cells were determined by RT- PCR,while the expression levels of LKB1,cyclin D1,Mcm2 and PCNA proteins by Western blot. Testing after transfection cell proliferation ability by CCK8. Results： Compared with those of blank and sh NC groups,the relative levels of LKB1,cyclin D1,Mcm2 and PCNAmRNAs and proteins in LKB1- shRNA1 cells increased significantly（ P〈 0. 01）. Conclusion： Gene silencing of LKB1 could up- regulate the expression of cyclin D1,Mcm2 and PCNA inprostate carcinoma cells; LKB1 gene might be a potential target for study on the molecular mechanism ofproliferation of prostatecarcinoma.

关 键 词：前列腺癌 LKB1基因 细胞周期素D1 微小染色体维持缺陷蛋白2 增殖细胞核抗原 

分 类 号：R735.2[医药卫生—肿瘤]