摘要
目的分析微小RNA(miRNA,miR)-6838-5p在卵巢癌组织中的表达,探讨miR-6838-5p是否通过靶向PPM1D抑制卵巢癌细胞的增殖和迁移。方法选取2018年3月至2019年11月本院39例卵巢癌组织及癌旁组织,实时荧光定量PCR(qRT-PCR)法检测组织中miR-6838-5p的表达水平。分别检测5种卵巢癌细胞株(OC3、SKOV-3、OVCAR-3、A2780、HO-8910)及正常卵巢上皮细胞(IOSE80)中miR-6838-5p的表达。选择miR-6838-5p表达水平最低的细胞株为对象,分为阴性对照组(转染NC模拟物)和miR-6838-5p组(转染miR-6838-5p模拟物)。细胞计数试剂盒(cell counting kit-8,CCK-8)和Transwell迁移实验分别检测各组细胞增殖和迁移能力。生物信息学技术和双荧光素酶报告基因实验分别预测和验证miR-6838-5p的靶基因。qRT-PCR和蛋白质印迹法(Western blot)检测各组细胞靶基因表达水平。结果miR-6838-5p在卵巢癌组织中相对表达水平低于癌旁组织,差异有统计学意义(P<0.01)。与正常卵巢上皮细胞相比,miR-6838-5p的表达水平在各卵巢癌细胞株降低(P<0.05),其中在OVCAR-3细胞中的表达最低(P<0.01)。过表达miR-6838-5p后,OVCAR-3细胞增殖能力在第2,3,4,5天低于阴性对照(P<0.05),OVCAR-3细胞迁移能力低于阴性对照(P<0.01)。生物信息学技术结合双荧光素酶报告基因实验显示miR-6838-5p的靶基因是PPM1D。过表达miR-6838-5p后,OVCAR-3细胞中PPM1D基因表达水平降低(P<0.01)。结论miR-6838-5p在卵巢癌组织和细胞株中的表达水平降低。miR-6838-5p可能通过靶向调控PPM1D基因的表达,抑制OVCAR-3细胞的增殖和迁移。
Objective To investigate the expression of microRNA(miR)-6838-5p in ovarian cancer tissues,and explore whether miR-6838-5p can inhibit the proliferation and the migration of ovarian cancer cells by targeting PPM1D.Methods A total of 39 pairs of ovarian cancer tissues and adjacent tissues in our hospital from March 2018 to November 2019 were selected to detect the expression level of miR-6838-5p by real-time fluorescence quantitative PCR(qRT-PCR)method.The expression levels of miR-6838-5p in five ovarian cancer cell lines(OC3,SKOV-3,OVCAR-3,A2780,HO-8910)and normal ovarian epithelial cells(IOSE80)were detected,respectively.Then the cell line with the lowest expression level of miR-6838-5p was selected as subjects and divided into negative control group(transfected with NC mimic)and miR-6838-5p group(transfected with miR-6838-5p mimic).Cell counting kit-8(CCK-8)and Transwell migration experiment were used to detect the cell proliferation and migration abilities,respectively.Bioinformatics technology and dual luciferase target experiment were used to respectively predict and verify the target gene of miR-6838-5p.QRT-PCR and Western blot were used to detect the expression levels of target genes in each group.Results The relative expression level of miR-6838-5p in ovarian cancer tissues was statistically lower than that in adjacent tissues(P<0.01).Compared with normal ovarian epithelial cells,the expression level of miR-6838-5p was reduced in each ovarian cancer cell line(P<0.05),and it was the lowest in OVCAR-3 cells(P<0.01).After overexpression of miR-6838-5p,the proliferation capacity of OVCAR-3 cells was lower in miR-6838-5p group than in negative control group at day 2,3,4,5(P<0.05),and the migration capacity of OVCAR-3 cells was lower than that of the negative control group(P<0.01).Bioinformatics technology combined with dual luciferase reporter gene experiments showed that the target gene of miR-6838-5p was PPM1D.Compared with negative control group,the PPM1D gene expression level was decreased in miR-6838-5p g
作者
胡翠芳
陈梦雨
黄寅虎
王婧
邱英
HU Cuifang;CHEN Mengyu;HUANG Yinhu;WANG Jing;QIU Ying(Department of Obstetrics and Gynecology,Daping Hospital,Army Medical University,Chongqing 400042,China)
出处
《山西医科大学学报》
CAS
2020年第12期1315-1320,共6页
Journal of Shanxi Medical University
