微小核糖核酸hsa-miR-663对肾上腺皮质细胞SW-13增殖及凋亡的影响  

Effect of hsa-miR-663 on proliferation and apoptosis of human adrenal cortical cell line SW-13

作　　者：赵欣[1] 张学斌[1] 李汉忠[1] 

机构地区：[1]中国医学科学院 北京协和医学院 北京协和医院泌尿外科 协和医学转化中心,100730

出　　处：《中华泌尿外科杂志》2016年第8期621-624,共4页Chinese Journal of Urology

基　　金：国家自然科学基金（81070628）

摘　　要：目的 在前期研究基础上,进一步探讨与促肾上腺皮质激素非依赖性肾上腺大结节增生（adrenocorticotropin-independent macronodular adrenal hyperplasia,AIMAH）发病相关的微小核糖核酸（microRNA）对肾上腺皮质细胞（SW-13细胞）增殖及凋亡的影响.方法 经前期筛选、验证研究发现AIMAH与正常肾上腺组织相比存在microRNA差异表达,其中hsa-miR-663表达明显上调.2015年10月至2016年2月,将hsa-miR-663作为目标microRNA,合成相应的microRNA mimics及inhibitors,转染SW-13细胞.通过实时定量逆转录聚合酶链反应（RT-PCR）确认hsa-miR-663在SW-13细胞中的转染效果.根据RT-PCR结果将经转染的细胞系分为5个组进行对比研究,分别为SW-13空白对照组、mimics阴性对照组、inhibitor阴性对照组、hsa-miR-663过表达组、hsa-miR-663敲低组,应用MTT法检测各组细胞的生长曲线,应用Annexin V-FITC/PI双染法检测各组细胞的凋亡.明确hsa-miR-663对SW-13细胞增殖及凋亡的影响.结果 采用RT-PCR确认,在mimics转染的细胞中,hsa-miR-663过表达;在Inhibitors转染的细胞中,hsa-miR-663表达敲低.MTT结果显示,与正常对照组及阴性对照组相比,hsa-miR-663过表达组细胞增殖明显,敲低组细胞增殖显著下降.Annexin V-FITC/PI双染法结果显示,与正常对照组及阴性对照组相比,hsa-miR-663过表达组细胞凋亡无明显变化,敲低组细胞凋亡轻度增加,差异无统计学意义（P＞0.05）.结论 hsa-miR-663过表达可以促进SW-13细胞增殖,可能与AIMAH发病相关.Objective To investigate the effect of has-miR-663 on proliferation and apoptosis of human adrenal cortical cell line SW-13.Methods Based on the first stage study,in adrenocorticotropinindependent macronodular adrenal hyperplasia （AIMAH）,hsa-miR-663 was up-regulated.The synthetic hsa-miR-663 mimics and inhibitors were transfected the SW-13 cell line.Real-time quantitative PCR （RT-PCR） tested the effect of the transfection.According to the results of RT-PCR,cell line was divided into five groups,including blank group,mimics NC group,inhibitor NC group,mimics group,inhibitor group.Then proliferating of the cell was tested by MTT and the apoptosis by Annexin V-FITC/PI.Result By RT-PCR,mimics group showed that has-miR-663 was over-expressed,and inhibitor group were knocked down.MTTshowed that,compared with NC group and Blank group,SW-13 cell line proliferated faster in mimics group and slower in inhibitor group.Annexin V-FITC/PI showed that apoptosis of SW-13 cell line was not different in all the five groups.Conclusion Up-regulating has-miR-663 could accelerate proliferation of adrenal cortical cells.The microRNA might play some role in the pathogenesis of AIMAH.

关 键 词：库欣综合征 促肾上腺皮质激素非依赖性肾上腺大结节增生 细胞增殖 细胞 凋亡 微小核糖核酸hsa—miR-663 

分 类 号：R586[医药卫生—内分泌]