circPRMT5通过靶向miR-98-5p对膀胱癌细胞增殖、迁移侵袭的机制研究  

Study on the mechanism of circPRMT5 targeting miR-98-5p on the proliferation,migration and invasion of bladder cancer cells

作　　者：谢奕彪[1] 姜良真 崔小健[1] XIE Yibiao;JIANG Liangzhen;CUI Xiaojian(Department of Urology,Tianjin Taida Hospital,Tianjin 300450,China)

机构地区：[1]天津市泰达医院泌尿外科,天津300450

出　　处：《中国实验诊断学》2024年第6期708-713,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探究circPRMT5通过靶向miR-98-5p对膀胱癌细胞增殖、迁移侵袭的机制研究。方法qRT-PCR检测永生化正常尿路上皮细胞SV-HUC-1和膀胱癌细胞(T24、J82、HT-1376)中circPRMT5、miR-98-5p和X染色体连锁的凋亡抑制蛋白(XIAP)mRNA相对表达水平。核糖核酸酶(RNase)R和放线菌素D的消化实验鉴定circPRMT5的环状结构。将T24细胞分为si-NC组、si-circPRMT5组、miR-NC组、miR-98-5p组、si-circPRMT5+anti-miR-NC组、si-circPRMT5+anti-miR-98-5p组。qRT-PCR检测circPRMT5和miR-98-5p表达水平;MTT、克隆实验检测细胞增殖水平;Transwell检测迁移、侵袭能力;Western Blot检测MMP2、MMP9、XIAP蛋白表达;双荧光素酶报告实验检测circPRMT5和miR-98-5p靶向关系。结果与永生化正常尿路上皮细胞SV-HUC-1相比,膀胱癌细胞T24、J82、HT-1376中circPRMT5、XIAP mRNA相对表达水平增加,miR-98-5p降低。circPRMT5比线性PRMT5稳定性更强。沉默circPRMT5或过表达miR-98-5p降低膀胱癌细胞存活率,减少克隆细胞数、迁移细胞数、侵袭细胞数,降低MMP2、MMP9、XIAP蛋白表达。circPRMT5靶向负调控miR-98-5p的表达,抑制miR-98-5p可部分回复沉默circPRMT5对膀胱癌细胞增殖、迁移、侵袭及XIAP mRNA和蛋白表达的作用。结论circPRMT5靶向负调控miR-98-5p抑制膀胱癌细胞增殖、迁移和侵袭能力。Objective To explore the mechanism of circPRMT5 targeting miR-98-5p on the proliferation, migration and invasion of bladder cancer cells.Methods qRT-PCR was used to detect the relative expression levels of circPRMT5,miR-98-5p and X-linked inhibitor of apoptosis(XIAP) mRNA in immortalized normal urothelial cells SV-HUC-1 and bladder cancer cells(T24,J82,HT-1376).The ring structure of circPRMT5 was identified by digestion experiments of ribonuclinase(RNase) R and actinomycin D.Divide T24 cells into si-NC group, si-circPRMT5 group, miR-NC group, miR-98-5p group, si-circPRMT5+anti-miR-NC group, si-circPRMT5+anti-miR-98-5p group.qRT-PCR to detect the expression levels of circPRMT5 and miR-98-5p;MTT,cloning experiments to detect cell proliferation;Transwell to detect migration and invasion ability;Western Blot to detect MMP2,MMP9 and XIAP protein expression;dual luciferase reporter experiment to detect the targeting relationship between circPRMT5 and miR-98-5p.Results Compared with the immortalized normal urothelial cells SV-HUC-1,the relative expression levels of circPRMT5 and XIAP mRNA in bladder cancer cells T24,J82,HT-1376 increased, and miR-98-5p decreased.circPRMT5 is more stable than linear PRMT5.Silencing circPRMT5 or overexpression of miR-98-5p reduces the survival rate of bladder cancer cells, reduces the number of cloned cells, the number of migrating cells, and the number of invasive cells, and reduces the protein expression of MMP2,MMP9 and XIAP.circPRMT5 targets and negatively regulates the expression of miR-98-5p, and inhibition of miR-98-5p can partially restore the effect of silencing circPRMT5 on the proliferation, migration, invasion and the expression of XIAP mRNA and protein of bladder cancer cells.Conclusion circPRMT5 targets and negatively regulates miR-98-5p to inhibit the proliferation, migration and invasion of bladder cancer cells.

关 键 词：circPRMT5 miR-98-5p 膀胱癌 增殖 迁移 侵袭 

分 类 号：R737.14[医药卫生—肿瘤]