miR-155-5p靶向YWHAZ影响22RV1细胞的增殖和迁移  

Proliferation and migration of 22RV1 cells affected by miR-155-5p targeting of YWHAZ

作　　者：李永 赵佳福[2] 许厚强[1,2] LI Yong;ZHAO Jia-fu;XU Hou-qiang(Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Collaborative Innovation Center for Mountain Ecology&Agro-Bioengineering(CICMEAB),Institute of Agro-bioengineering/College of Life Sciences,Guizhou University,Guiyang 550025,China;Key Laboratory of Plateau Mountain Animal Genetics,Breeding and Reproduction,Ministry of Education,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学农业生物工程研究院/生命科学学院,山地植物资源保护与种质创新教育部重点实验室,山地生态与农业生物工程协同创新中心,贵州贵阳550025 [2]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室,贵州贵阳550025

出　　处：《生物技术》2022年第4期432-437,共6页Biotechnology

基　　金：国家自然科学基金项目(31860242);贵州省科学技术基金项目(黔科合基础[2020]1Y085)。

摘　　要：[目的]研究miR-155-5p是否靶向YWHAZ调控22RV1细胞的增殖和迁移。[方法]数据库筛选靶向YWHAZ的micRNA;过表达miR-155-5p后用RT-qPCR检测YWHAZ表达水平;CCK-8和划痕试验检测miR-155-5p是否靶向调控YWHAZ影响22RV1的增殖和迁移。在线数据库筛选miR-155-5p与YWHAZ 3'UTR的结合靶点,通过将YWHAZ的3'UTR插入双荧光素酶报告系统中进行互作位点的检测。[结果]miR-155-5p在多个数据库中均靶向YWHAZ且得分最高,且miR-155-5p的过表达,极显著抑制了YWHAZ的表达(P<0.01);CCK-8和细胞划痕结果表明,转染miR-155-5p的试验组,22RV1细胞增殖和迁移能力均显著低于NC组(P<0.05)。双荧光素酶试验结果表明转染miR-155-5p+pmriGLO-YWHAZ后22RV1荧光活性极显著低于转染NC+pmriGLO-YWHAZ组(P<0.01),说明miR-155-5p靶向结合YWHAZ的3'UTR区。[结论]miR-155-5p靶向结合YWHAZ的3'UTR区,极显著抑制了YWHAZ的表达(P<0.01)。最终显著抑制了22RV1细胞增殖和迁移能力(P<0.05),相关结果为研究前列腺癌症靶向药物提供理论基础和实验思路。[Objective]To investigate whether miR-155-5p targets YWHAZ to regulate the proliferation and migration of 22RV1 cells.[Method]Database screening for micRNAs targeting YWHAZ.Detection of YWHAZ expression levels by RT-qPCR after overexpression of miR-155-5p.CCK-8 and scratch assays test whether miR-155-5p targets and regulates YWHAZ to affect the proliferation and migration of 22RV1.Online database screening of miR-155-5p binding target to YWHAZ 3'UTR by inserting the 3'UTR of YWHAZ into a dual luciferase reporter system for the detection of reciprocal sites.[Result]Revealed that miR-155-5p targeted YWHAZ and scored highest in several databases,and overexpression of miR-155-5p inhibited YWHAZ expression highly significantly(P<0.01);CCK-8 and cell scratch results showed that the proliferation and migration ability of 22RV1 cells were significantly lower in the test group transfected with miR-155-5p than in the NC group(P<0.05).The results of dual luciferase assay showed that the fluorescence activity of 22RV1 after transfected with miR-155-5p+pmriGLO-YWHAZ was extremely significantly lower than that of the group transfected with NC+pmriGLO-YWHAZ(P<0.01),indicating that miR-155-5p targets binding to the 3'UTR region of YWHAZ.[Conclusion]miR-155-5p targeting binding to the 3'UTR region of YWHAZ inhibited the expression of YWHAZ extremely significantly(P<0.01).Finally,it significantly inhibited the proliferation and migration ability of 22RV1 cells(P<0.05),and the related results provide theoretical basis and experimental ideas for the study of prostate cancer targeting drugs.

关 键 词：前列腺癌 micoRNA 实时荧光定量PCR 双荧光素酶 细胞增殖 细胞迁移 

分 类 号：R737.25[医药卫生—肿瘤]