沉默TK1对前列腺癌PC3细胞增殖及侵袭的影响  

Silencing TK1 gene inhibits the proliferation and invasion of prostate cancer PC3 cells

作　　者：吕磊[1] 黄遂斌[1] 吴维[1] 柳懿鹏[1] 蒋国松[2] 周高峰[1] LYU Lei;HUANG Sui-bin;WU Wei;LIU Yi-peng;JIANG Guo-song;ZHOU Gao-feng(Department of Urology,Wuhan First Hospital,Wuhan 430022,China;Department of Urology,Union Hospital,Huazhong University of Science and Technology,Wuhan 430022,China)

机构地区：[1]武汉市第一医院泌尿外科,湖北武汉430022 [2]华中科技大学附属协和医院泌尿外科,湖北武汉430022

出　　处：《中华肿瘤防治杂志》2021年第19期1456-1461,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81502204,81974396);湖北省自然科学基金(2020CFB732);武汉市科技计划(2020020601012325)。

摘　　要：目的探讨胸苷激酶-1(TK1)对前列腺癌PC3细胞增殖和侵袭能力的影响。方法实时荧光定量PCR和蛋白质印迹法检测TK1mRNA和蛋白在人正常前列腺上皮细胞和前列腺癌PC3细胞中的表达;利用脂质体转染PC3细胞,根据转染情况将实验分为对照组(未转染siRNA)、NC siRNA(转染阴性siRNA)组和TK1siRNA组(转染TK1siRNA)。转染48h后,通过MTT细胞增殖实验、克隆形成实验和侵袭实验检测各组PC3细胞的存活能力、增殖活性和侵袭活性。结果人正常前列腺上皮细胞RWPE-1和前列腺癌PC3细胞中TK1mRNA相对表达水平分别为1.000±0.022和6.512±0.708,组间均值差异有统计学意义,t=13.612,P=0.005;TK1蛋白相对表达水平分别为1.000±0.031和3.508±0.583,组间均值差异有统计学意义,t=20.663,P=0.002。PC3细胞转染NC siRNA和TK1siRNA 48h后,TK1mRNA在对照组中的相对表达量为1.000±0.013,NC siRNA组为1.023±0.018,TK1siRNA组为0.142±0.050,3组比较差异有统计学意义,F=44.27,P<0.001。其中,TK1siRNA组TK1mRNA表达量显著低于对照组(t=14.625,P=0.005)和NC siRNA组(t=13.374,P=0.006),差异有统计学意义。TK1蛋白在对照组中的相对表达量为1.000±0.041,NC siRNA组为1.018±0.052,TK1siRNA组为0.250±0.062,3组比较差异有统计学意义,F=42.52,P<0.001。其中,TK1siRNA组TK1蛋白表达量显著低于对照组(t=21.330,P=0.002)和NC siRNA组(t=16.426,P=0.004),差异有统计学意义。MTT结果显示,TK1siRNA组PC3细胞的存活能力降低,与NC siRNA组比较差异有统计学意义,并呈时间依赖,F_(组别)=68.37,F_(时间)=114.88,F_(组别×时间)=43.62,均P<0.001。克隆形成实验结果显示,NC siRNA组和TK1siRNA组克隆细胞集落数分别为32.25±5.82和8.64±1.50,2组比较差异有统计学意义,t=32.181,P=0.001。细胞侵袭实验结果显示,NC siRNA组和TK1siRNA组的侵袭细胞数分别为158.35±18.62和60.52±7.33,2组比较差异有统计学意义,t=28.742,P=0.001。结论TK1mRNA和蛋白在前列腺癌PC3细胞中呈Objective To investigate the effect of TK1on proliferation and invasion of prostate cancer PC3cells in vitro.Methods The expression levels of TK1mRNA and protein in human normal prostate epithelial cells RWPE-1and prostate cancer cells PC3 were determined by quantitative real time PCR(qRT-PCR)and western blot,respectively.After transfecting with NC siRNA or TK1siRNA for 48h,the cell viability,proliferation activity and invasion activity of PC3cells were determined by MTT assay,clone formation assay and invasion assay.Results The relative expression levels of TK1mRNA in human normal prostate epithelial cells RWPE-1and prostate cancer cells PC3were 1.000±0.022and6.512±0.708,respectively,and the difference was statistically significant(t=13.612,P=0.005).The relative expression levels of TK1protein in human normal prostate epithelial cells RWPE-1and prostate cancer cells PC3were 1.000±0.031and 3.508±0.583,respectively,and the difference was statistically significant(t=20.663,P=0.002).After transfection with NC siRNA or TK1siRNA for 48h,the relative mRNA levels of TK1in control group,NC siRNA group and TK1siRNA group were 1.000±0.013,1.023±0.018and 0.142±0.050,respectively,and the difference was statistically significant(F=44.27,P<0.001).Among them,the relative expression levels TK1mRNA in TK1siRNA group was significantly lower than that in control group(t=14.625,P=0.005)and NC siRNA group(t=13.374,P=0.006).The relative protein levels of TK1of PC3cells in control group,NC siRNA group and TK1siRNA group were 1.000±0.041,1.018±0.052and 0.250±0.062,respectively,and the difference was statistically significant(F=42.52,P<0.001).Among them,the relative expression levels TK1protein in TK1siRNA group was significantly lower than that in control group(t=21.330,P=0.002)and NC siRNA group(t=16.426,P=0.004).MTT assay showed that the cell viability of PC3cells decreased in TK1siRNA group,and the difference was statistically significant,compared with the NC siRNA group,and it was time-dependent,F_(group)=68.37,F_(time)=11

关 键 词：胸苷激酶-1 前列腺癌 增殖 侵袭 

分 类 号：R737.25[医药卫生—肿瘤]