RNA干扰LKB1基因对前列腺癌细胞转化生长因子-β1表达的影响  

Effect of silencing LKB1 gene by shRNA on expression of transforming growth factor-β1 in prostate cancer cells

作　　者：邓玮[1] 赵世巧[2] 陈潇[3] 刘斌[4] 吕玉宇[1] DENG We;ZHAO Shi-qiao;CHEN Xiao;LIU Bin;LV Yu-yu(Department of Pathology, The Fifth People＇s Hospital of Chongqing City, Chongqing 400062, Chin)

机构地区：[1]重庆市第五人民医院病理科,重庆400062 [2]重庆市中医院检验科,重庆400011 [3]重庆市沙坪坝区人民医院康复科,重庆400030 [4]重庆医科大学基础医学院病理教研室,重庆400016

出　　处：《中国生物制品学杂志》2018年第6期598-602,共5页Chinese Journal of Biologicals

基　　金：重庆市南岸区科委资助项目(2011年第二批;项目序号:28)

摘　　要：目的探讨沉默肝激酶B1/丝氨酸-苏氨酸激酶11(liver kinase B1/serine-threonine kinase 11,LKB1/STK11),基因对前列腺癌细胞株RWPE-1中转化生长因子-β1(transforming growth factor-β1,TGF-β1)的调控作用。方法构建3个sh RNA LKB1重组载体,转染至高表达LKB1的前列腺癌细胞株RWPE-1中,RT-PCR、Western blot法分别检测LKB1及TGF-β1 m RNA及蛋白表达水平;ELISA法检测细胞培养液上清中TGF-β1的分泌水平。结果构建获得的3种sh RNA LKB1表达载体,均能抑制LKB1基因的表达,其中以LKB1-sh RNA1沉默效果最强,其对LKB1 m RNA水平的抑制率为74.20%,对蛋白水平的抑制率为71.66%;转染重组质粒LKB1-sh RNA1后,TGF-β1的m RNA、蛋白表达及分泌水平均明显增加(P﹤0.05)。结论 LKB1基因沉默后,TGF-β1的表达明显增高,LKB1参与调节前列腺癌细胞中TGF-β1的表达,提示LKB1基因可作为研究前列腺癌发生发展的分子机制的新靶点。Objective To investigate the regulatory effect of silencing liver kinase B1/serine-threonine kinase 11（LKB1/STK11） gene on transforming growth factor-β1（TGF-β1） in prostate cancer cells. Methods Three specific sh RNA plasmids were constructed based on LKB1 sequence and transfected to prostate carcinoma RWPE-1 cells highly expressing LKB1. The expression levels of LKB1 and TGF-β1 m RNAs were determined by RT-PCR,while those of the two proteins by Western blot. The secretion level of TGF-β1 in cell culture supernatant was determined by ELISA.Results Three LKB1 sh RNA expression plasmids were successfully constructed,which inhibited the expression of LKB1 gene. Of the plasmids,LKB1-sh RNA1 showed the strongest inhibitory effect,by which the inhibitory rate of LKB1 m RNA expression was 74. 20% while that of protein expression was 71. 66%. The m RNA and protein expressions and secretion levels of TGF-β1 increased significantly in the cells transfected with LKB1-sh RNA1（P〈0. 05）. Conclusion The expression level of TGF-β1 increased significantly after silencing of LKB1 gene,indicating that LKB1 possibly participle in regulation of TGF-β1 expression in prostate cancer cells. These data suggest that LKB1 gene may be used as a potential target for exploring the molecular mechanism of tumorgenesis of gastric cancer.

关 键 词：前列腺癌 丝氨酸-苏氨酸激酶11 转化生长因子-Β1 RNA干扰 

分 类 号：R737.2[医药卫生—肿瘤]