敲低WWP1对肺癌SPC-A1细胞增殖和侵袭能力影响及其机制  

Effect of knockdown of WWP1 on the proliferation and invasion of lung cancer SPC-A1 cells and its mechanism

作　　者：张成伟 毕昕 闫睿 刘艳超 ZHANG Cheng-wei;BI Xin;YAN Rui;LIU Yan-chao(Department of Thoracic Surgery,Electric Power Teaching Hospital,Capital Medical University,Beijing 100073,China;Department of Thoracic Surgery,Third Medical Center,Chinese PLA General Hospital,Beijing 100039,China)

机构地区：[1]首都医科大学电力教学医院胸外科,北京100073 [2]解放军总医院第三医学中心胸外科,北京100039

出　　处：《中华肿瘤防治杂志》2021年第16期1203-1208,共6页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的检测肺癌细胞中WW结构域E3泛素蛋白连接酶1(WWP1)和人第10号染色体缺失的磷酸酶(PTEN)表达,并探讨敲低WWP1表达对肺癌SPC-A1细胞增殖和侵袭能力影响机制。方法利用qRT-PCR和蛋白质印迹法检测肺癌细胞SPC-A1、H1299及正常肺上皮细胞BEAS-2B中WWP1和PTEN表达;WWP1 siRNA转染SPC-A1肺癌细胞以敲低WWP1表达,分为si-NC组,siWWP1#1组及siWWP1#2组,利用qRT-PCR和蛋白质印迹法检测各组细胞WWP1和PTEN表达,通过Transwell法检测肺癌细胞迁移和侵袭,细胞计数试剂(CCK-8)法检测细胞增殖。结果在正常肺上皮细胞BEAS-2B,肺癌细胞SPC-A1和H1299中WWP1 mRNA相对表达量分别为1.00±0.08、3.10±0.24和2.30±0.20,差异有统计学意义,F=93.960,P<0.001;PTEN表达量分别为1.00±0.10、0.34±0.04和0.53±0.04,差异有统计学意义,F=91.940,P<0.001;与正常细胞株相比,WWP1在肺癌细胞株中表达增加,而PTEN表达则降低。WWP1和PTEN蛋白检测结果与qRT-PCR检测结果一致。敲低WWP1后,si-NC组、siWWP1#1组和siWWP1#2组WWP1 mRNA相对表达量分别为1.00±0.09、0.32±0.03和0.27±0.03,差异有统计学意义,F=151.200,P<0.001;PTEN表达量分别为1.00±0.07、2.81±0.21和3.34±0.33,差异有统计学意义,F=85.810,P<0.001,与si-NC组相比,WWP1在siWWP1#1组,siWWP1#2组表达降低,而PTEN表达则增加。WWP1和PTEN蛋白检测结果与qRT-PCR检测结果一致。Transwell迁移结果显示,si-NC组、siWWP1#1组和siWWP1#2组细胞迁移数目分别为(126±9)、(44±4)和(50±4)个,差异有统计学意义,F=166.400,P<0.001;Transwell侵袭结果显示,si-NC组、siWWP1#1组和siWWP1#2组细胞侵袭数目分别为(104±10)、(40±3)和(46±5)个,差异有统计学意义,F=83.910,P<0.001;与si-NC组相比,siWWP1#1组,siWWP1#2组细胞迁移和侵袭数目均降低。CCK-8法检测敲低WWP1对肺癌细胞增殖活力影响结果显示,si-NC组,siWWP1#1组,siWWP1#2组细胞增殖活力分别为(100±8)%、(52±4)%和(42±3)%,差异有统计学意义,F=173.200,P<0.001�Objective To detect the expression of WWP1 and PTEN in lung cancer cells, and explore the mechanism of knocking down WWP1 expression on the proliferation and invasion of lung cancer SPC-A1 cells.Methods qRT-PCR and western blotting were used to detect the expression of WWP1 and PTEN in lung cancer cells SPC-A1,H1299 and normal lung epithelial cells BEAS-2 B.WWP1 siRNA transfected SPC-A1 lung cancer cells to knock down the expression of WWP1 and divided into si-NC group, siWWP1#1 group and siWWP1#2 group.The expressions of WWP1 and PTEN of each group were detected by qRT-PCR and western blotting.The migration and invasion of lung cancer cells were detected by Transwell method.Cell counting reagent(CCK-8) method was used to detect cell proliferation.Results The relative expression of WWP1 mRNA in normal lung epithelial cells BEAS-2 B,lung cancer cells SPC-A1 and H1299 were 1.00±0.08,3.10±0.24 and 2.30±0.20,respectively,the difference was statistically significant,F=93.960,P<0.001.The expression levels of PTEN were 1.00±0.10,0.34±0.04 and 0.53±0.04,respectively,the difference was statistically significant,F=91.940,P<0.001.Compared with normal cell lines,WWP1 expression increased in lung cancer cell lines,while PTEN expression showed a decrease.The WWP1 and PTEN protein detection results were consistent with the qRT-PCR detection results.After knocking down WWP1,the relative expression of WWP1 mRNA in the si-NC group,siWWP1#1 group and siWWP1#2 group were 1.00±0.09,0.32±0.03 and 0.27±0.03,respectively,the difference was statistically significant,F=151.200,P<0.001.PTEN expression levels were 1.00±0.07,2.81±0.21 and 3.34±0.33,respectively,the difference was statistically significant,F=85.810,P<0.001.Compared with si-NC group,expression of WWP1 in siWWP1#1 group,siWWP1#2 group was found to decrease,while PTEN expression was increased.The WWP1 and PTEN protein detection results were consistent with the qRT-PCR detection results.Transwell migration results showed that the number of cell migration in the si-NC

关 键 词：肺癌 WW结构域E3泛素蛋白连接酶1 人第10号染色体缺失的磷酸酶 增殖 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]