miR-98-5p靶向PYGO2基因对宫颈癌细胞增殖、迁移和侵袭的影响及其作用机制研究  

Effect and mechanisms of miR-98-5p targeting PYGO2 gene on proliferation,migration and invasion of cervical cancer

作　　者：首泉[1] 王茜 谭志远 Shou Quan;Wang Qian;Tan Zhiyuan(Department of Gynecology,the First People's Hospital of Chenzhou City,Chenzhou 423000,China;Department of Anesthesiology,Chenzhou Fourth People's Hospital,Chenzhou 423000,China)

机构地区：[1]湖南省郴州市第一人民医院妇科,郴州423000 [2]湖南省郴州市第四人民医院麻醉科,郴州423000

出　　处：《中华细胞与干细胞杂志（电子版）》2020年第3期138-145,共8页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的探讨miR-98-5p对宫颈癌细胞增殖、迁移和侵袭的影响以及其作用机制。方法选取2016年1月至2019年1月郴州市第一人民医院收治的宫颈癌患者的癌组织和癌旁组织;予以miR-98-5p、si-PYGO2及anti-miR-98-5p单独或共培养Siha细胞,记为:miR-NC组、miR-98-5p组、si-NC组、si-PYGO2组、anti-miR-NC组、anti-miR-98-5p组、miR-98-5p+pcDNA组、miR-98-5p+pcDNA-PYGO2组。运用qRT-PCR检测宫颈癌组织和细胞中miR-98-5p和PYGO2 mRNA的表达水平;Western blot检测蛋白表达;MTT法检测细胞增殖活性;Transwell检测细胞迁移和侵袭;将WT-PYGO2、MUT-PYGO2分别与miR-NC、miR-98-5p共转染至Siha细胞中,双荧光素酶报告基因检测实验检测荧光活性。采用方差分析和t检验进行统计学分析。结果与癌旁组织相比,宫颈癌组织中miR-98-5p表达水平降低(0.98±0.08比0.47±0.05),PYGO2 mRNA(1.00±0.07比2.43±0.24)和蛋白表达水平(0.27±0.03比0.62±0.05)均升高(P均<0.001)。与正常宫颈细胞Ect1/E6E7相比,宫颈癌细胞Siha、Hela、Caski中PYGO2 mRNA(0.98±0.09比2.76±0.23、2.46±0.24、2.55±0.21)和蛋白表达水平(0.21±0.03比0.62±0.06、0.51±0.05、0.57±0.06)升高;miR-98-5p的表达水平降低(1.00±0.08比0.34±0.04、0.56±0.05、0.46±0.04)(P均<0.05)。与miR-NC组相比,miR-98-5p组宫颈癌Siha细胞活性(48 h:0.61±0.05比0.42±0.04,72 h:1.02±0.09比0.59±0.06)、迁移数量[(112.46±10.27)个比(48.35±4.96)个]及侵袭数量[(92.47±9.56)个比(39.46±3.52)个]均降低(P均<0.05)。与si-NC组相比,si-PYGO2组宫颈癌Siha细胞活性(48 h:0.64±0.06比0.46±0.05,72 h:1.05±0.08比0.67±0.06)、Siha迁移数量[(106.48±9.75)个比(42.16±4.25)个]和侵袭数量[(87.63±8.11)个比(35.42±6.20)个]均降低(P均<0.05);Cyclin D1、MMP-2、MMP-9、MMP-14表达水平降低,p21、p27表达水平升高,差异有统计学意义(P均<0.05)。与miR-NC组比较,miR-98-5p组转染WT-PYGO2的Siha细胞荧光素酶活性(0.38±0.04比0.99±0.08)降低(P<0.05),转染MUT-PYGOObjective To investigate the effects and mechanisms of miR-98-5p on the proliferation,migration and invasion of cervical cancer cells.Methods Cancer tissues and adjacent tissues of cervical cancer patients admitted to the first people's Hospital of Chenzhou City from January 2016 to January 2019 were collected;miR-98-5p,si-PYGO2 and anti-miR-98-5p were individually or co-cultured in Siha cells,and named miR-NC group,miR-98-5p group,and si-NC Group,si-PYGO2 group,anti-miR-NC group,anti-miR-98-5p group,miR-98-5p+pcDNA group,miR-98-5p+pcDNA-PYGO2 group.qRT-PCR was used to detect the expressions of miR-98-5p and PYGO2 mRNA in cervical cancer tissues and cells;Western blot was used to detect protein expression;MTT method to detect cell proliferation activity;Transwell to detect cell migration and invasion.WT-PYGO2 and MUT-PYGO2 were co-transfected with miR-NC and miR-98-5p into Siha cells,and the fluorescent activity was detected by dual luciferase reporter gene detection experiment.Statistical analysis was performed using method analysis and t-test.Results Compared with adjacent tissues,the expression of miR-98-5p in cervical cancer tissue was significantly reduced(0.98±0.08 vs 0.47±0.05),PYGO2 mRNA(1.00±0.07 vs 2.43±0.24)and protein expression(0.27±0.03 vs 0.62±0.05)was increased(all P<0.001).Compared with normal cervical cells Ect1/E6E7,PYGO2 mRNA(0.98±0.09 vs 2.76±0.23,2.46±0.24,2.55±0.21)and protein levels(0.21±0.03 vs 0.62±0.06,0.51±0.05,0.57±0.06)in cervical cancer cells Siha,Hela,and Caski was increased;the expression of miR-98-5p(1.00±0.08 vs 0.34±0.04,0.56±0.05,0.46±0.04)was decreased(all P<0.05).Compared with the miR-NC group,the activity of cervical cancer Siha cells in the miR-98-5p group(48 h:0.42±0.04 vs 0.61±0.05,72 h:0.59±0.06 vs 1.02±0.09),the number of migration(48.35±4.96 vs 112.46±10.27)and invasions(39.46±3.52 vs 92.47±9.56)were decreased(all P<0.05).Compared with the si-NC group,the activity of cervical cancer Siha cells in the si-PYGO2 group(48 h:0.46±0.05 vs 0.64±0.

关 键 词：miR-98-5p PYGO2 宫颈癌 增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]