摘要
目的:探讨微小RNA(miR)-424靶向神经纤毛蛋白2(NRP2)对宫颈癌SiHa细胞增殖、迁移及顺铂敏感性的影响。方法:实时荧光定量PCR法和免疫印迹法检测宫颈癌组织和癌旁组织以及宫颈癌细胞SiHa和人宫颈上皮细胞HCerEpiC中miR-424和NRP2蛋白表达情况。将体外培养的SiHa细胞分为对照组、miR-ctrl组(转染miR-ctrl)、miR-424组(转染miR-424模拟物)、miR-424+pcDNA3.1组(共转染miR-424模拟物和pcDNA3.1空载体质粒)和miR-424+NRP2组(共转染miR-424模拟物和pcDNA3.1-NRP2过表达载体质粒),采用实时荧光定量PCR法验证转染效率,免疫印迹法检测SiHa细胞中NRP2蛋白表达,双荧光素酶报告基因实验检测miR-424和NRP2靶向关系,MTT法检测细胞增殖和顺铂敏感性,Transwell小室检测细胞迁移。结果:双荧光素酶报告基因实验证实miR-424可与NRP2靶向结合;相较于癌旁组织,宫颈癌组织中miR-424表达明显降低,NRP2蛋白表达明显升高(P<0.05);相较于HCerEpiC细胞,SiHa细胞中miR-424表达明显降低,NRP2蛋白表达明显升高(P<0.05);与miR-ctrl组比较,miR-424组SiHa细胞中miR-424表达水平明显升高,而NRP2蛋白表达水平、细胞增殖活力、迁移能力和顺铂对细胞的半数抑制浓度(IC50)值均明显降低(P<0.05);与miR-424+pcDNA3.1组比较,miR-424+NRP2组细胞中NRP2蛋白表达水平、细胞增殖活力、迁移能力和顺铂对细胞的IC50值均明显升高(P<0.05);miR-ctrl组和对照组之间以及miR-424+pcDNA3.1组和miR-424组之间上述各指标差异均无统计学意义(P>0.05)。结论:miR-424可通过靶向调控NRP2表达抑制SiHa细胞增殖、迁移并增强顺铂敏感性。
Objective:To investigate the effects of microRNA(miR)-424 targeting neuropilin-2(NRP2)on the proliferation,migration and cisplatin sensitivity of cervical cancer SiHa cells.Methods:The expression of miR-424 and NRP2 protein in cervical cancer tissues,para-cancerous tissues,cervical cancer cells SiHa and human cervical epithelial cells were detected by real-time fluorescent quantitative PCR and Western blotting.SiHa cells cultured in vitro were divided into control group,miR-ctrl group(transfected with miR-ctrl),miR-424 group(transfected with miR-424 mimic),miR-424+pcDNA3.1 group(co transfection of miR-424 mimic and pcDNA3.1 empty vector plasmid)and miR-424+NRP2 group(co transfection of miR-424 mimic and pcDNA3.1-NRP2 overexpression vector plasmid).Real-time fluorescence quantitative PCR was used to verify the transfection efficiency.The expression of NRP2 protein in SiHa cells was detected by Western blot,double luciferase reporter gene assay was used to detect the targeting relationship between miR-424 and NRP2,MTT assay was used to detect the proliferation and cisplatin sensitivity of SiHa cells,and Transwell chamber was used to detect the migration of SiHa cells.Results:Double luciferase reporter gene assay confirmed that miR-424 could targetingly bind to NRP2.Compared with para-cancerous tissues,the expression of miR-424 in cervical cancer tissues was significantly decreased,and the expression of NRP2 protein was significantly increased(P<0.05).Compared with those in miR-ctrl group,the expression level of miR-424 in SiHa cells in miR-424 group was significantly higher,while the protein expression level of NRP2,cell proliferation activity,migration ability and cisplatin half inhibitory concentration(IC50)value were significantly lower(P<0.05).Compared with those in miR-424+pcDNA3.1 group,the protein expression level of NRP2,cell proliferation activity,migration ability and IC50 value were significantly higher in miR-424+NRP2 group(P<0.05).While there was no significant difference in the above indexes between m
作者
付小玲
郭哲
许静
唐旭
曾辉
史惠蓉
FU Xiaoling;GUO Zhe;XU Jing;TANG Xu;ZENG Hui;SHI Huirong(Department of Gynecology,Nanyang Central Hospital,Henan Nanyang 473000,China;Department of Gynecology,the First Affiliated Hospital of Zhengzhou University,Henan Zhengzhou 450000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第10期1741-1747,共7页
Journal of Modern Oncology
