丝氨酸-苏氨酸激酶11基因沉默对人前列腺癌RWPE-1细胞侵袭及迁移能力的影响及其作用机制

Effects of liver kinase B1/serine-threonine kinase 11 gene silence on invasion and migration abilities of human prostate cancer RWPE-1 cells and relevant mechanism

目的探讨丝氨酸-苏氨酸激酶11(liver kinase B1/serine-threonine kinase 11,LKB1)基因沉默对人前列腺癌RWPE-1细胞侵袭及迁移能力的影响及其作用机制。方法将质粒LKB1-sh RNA1及sh NC分别转染至RWPE-1细胞中,同时设空白对照组(未转染),经G418筛选出稳定表达细胞株。RT-PCR及Western blot法检测RWPE-1细胞中LKB1基因mRNA转录及蛋白表达水平;划痕试验及Transwell小室试验分别检测LKB1基因沉默对RWPE-1细胞迁移及侵袭能力的影响;RT-PCR及Western blot法检测RWPE-1细胞中转化生长因子-β1(transforming growth factor-β1,TGF-β1)、基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及MMP-9基因mRNA转录及蛋白的表达水平。结果与sh NC组及空白对照组比较,LKB1-shRNA1组RWPE-1细胞中LKB1基因mRNA转录及蛋白表达水平明显降低(P<0.001);细胞划痕愈合能力及侵袭细胞数明显升高(P<0.001);TGF-β1、MMP-2及MMP-9基因mRNA转录及蛋白表达水平均明显上升(P<0.01)。结论 LKB1基因沉默可显著提高RWPE-1细胞的转移及侵袭能力,可能是通过上调TGF-β1、MMP-2及MMP-9表达水平产生作用的。

Objective To investigate the effects of liver kinase B1/serine-threonine kinase 11（LKB1） gene silence on invasion and migration abilities of human prostate cancer RWPE-1 cells as well as the relevant mechanism. Methods RWPE-1 cells were transfected with plasmids LKB1-sh RNA1 and sh NC respectively,using those untransfected as blank control,from which those stably expressing LKB1 were screened with G418. The LKB1 mRNA transcription and protein expression levels of LKB1 in RWPE-1 cells were determined by RT-PCR and Western blot respectively. The effects of LKB1 gene silence on invasion and migration abilities of RWPE-1 cells were evaluated by wound healing test and Transwell assay respectively. The mRNA transcription and protein expression levels of transforming growth factor-β1（TGF-β1）,matrix metalloproteinase-2（MMP-2）and MMP-9 were determined by RT-PCR and Western blot respectively.Results As compared with those in sh NC and blank control groups,the mRNA transcription and protein expression levels of LKB1 in LKB1-shRNA1 group decreased（P〈0. 001）,while the healing ability and number of transmembrane cells（P〈0. 001）as well as the mRNA transcription and protein expression levels of TGF-β1,MMP-2 and MMP-9（P〈0. 01）increased significantly. Conclusion LKB1 gene silence increased the migration and invasion abilities of RWPE-1 cells by a possible mechanism of up-regulating the expression levels of TGF-β1,MMP-9 and MMP-2.