miR-21对视网膜母细胞瘤细胞增殖的影响及作用机制  

Effects of MiR-21 on the proliferation of retinoblastoma and its mechanism

作　　者：桂馥[1] 吴宏禧[1] 游志鹏[1] 章余兰[1] GUI Fu;WU Hongxi;YOU Zhipeng;ZHANG Yulan(Department of Ophthalmology,the Second Affiliated Hospital of Nanchang University,Nanchang 330006,Jiangxi Province,China)

机构地区：[1]南昌大学第二附属医院眼科,江西省南昌市330006

出　　处：《眼科新进展》2020年第7期630-633,共4页Recent Advances in Ophthalmology

摘　　要：目的探讨MicroRNA-21(miR-21)对视网膜母细胞瘤(RB)细胞增殖的影响及作用机制。方法采用Real-time PCR技术检测miR-21在正常视网膜组织和确诊RB组织中的表达情况;然后在转染的基础上运用MTT检查RB细胞存活率、流式细胞仪检测细胞凋亡率。Western blot法检测与细胞凋亡相关蛋白PDCD4、Bax、Bcl-2的表达。结果与正常视网膜组织miR-21表达(0.703±0.071)相比,RB组织miR-21为高表达(2.214±0.162),差异有统计学意义(P<0.01)。在Weri-Rb-1细胞中,与NC组(2.245±0.213)相比,miR-21抑制剂转染后明显降低了miR-21的表达水平,miR-21 inhibitor组为0.683±0.075,差异有统计学意义(P<0.01)。两组细胞转染后24 h、48 h、72 h、96 h,MTT测定法检测细胞活力结果显示两组24 h的A值比较,差异无统计学意义(P>0.05),miR-21 inhibitor组在48 h、72 h、96 h的A值均低于NC组,差异均有统计学意义(均为P<0.01)。流式细胞术检测结果显示NC组凋亡细胞在总细胞中百分比为(3.045±0.301)%和(4.832±0.493)%,miR-21 inhibitor组凋亡细胞在总细胞中百分比为(2.593±0.257)%和(40.167±4.014)%,miR-21 inhibitor组Weri-Rb-1细胞的凋亡率明显高于miR-21 NC组(P<0.01)。Western blot检测结果显示NC组PDCD4表达(0.192±0.045)相比miR-21 inhibitor组(0.683±0.091)表达明显减少,NC组Bax的蛋白表达水平(0.143±0.036)相比miR-21 inhibitor组(1.192±0.054)也明显减少,差异均有统计学意义(P<0.01),NC组Bcl-2蛋白表达(0.864±0.038)相比miR-21 inhibitor组(0.257±0.026)明显增多,差异有统计学意义(P<0.05)。结论miR-21是RB的促癌基因,miR-21抑制剂可以通过降低miR-21表达抑制肿瘤细胞的增殖、促进细胞凋亡,这一过程与PDCD4、Bax、Bcl-2等凋亡相关蛋白有关。Objective To investigate the effect of MicroRNA-21(miRNA-21)on retinoblastoma(RB)cell proliferation and its possible mechanisms.Methods Expression of miR-21 in RB tissue and human normal tissue was measured Real-time PCR.RB cells were transfected with miR-21 control or miR-21 inhibitor via LipofectaminTM 2000.Cell viability was analyzed by MTT assay.Flow cytometry with Annexin V-FITC/PI reagent was used to detect cell apoptosis.The protein levels of Bax,Bcl-2and PDCD4 were examined by Western blot.Results miR-21 was overexpressed in RB tissues(2.214±0.162)when compared with normal retinal tissues(0.703±0.071)with significant difference(P<0.01).When compared with normal control(NC)group(2.245±0.213),in Weri-Rb-1 cells,miR-21 inhibitor transfection suppressed the expression of miR-21 in miR-21 inhibitor group(0.683±0.073)(P<0.01).In addition,24 hours,48 hours,72 hours,96 hours after transfection of the two groups of cells,the results of the viability of the cells via MTT assay showed that there was no statistically significant difference in the A value between the two groups at 24 hours(P>0.05),A values of miR-21 inhibitor group at 48 h,72 h,96 h were lower than those of the NC group,and the differences are statistically significant(all P<0.01).The results of flow cytometry showed that the percentage of apoptotic cells in the NC group was(3.045±0.301)%and(4.832±0.493)%,and(2.593±0.257)%and(40.167±4.014)%in the miR-21 inhibitor group.The apoptosis rate of the Weir-Rb-1 cells in the miR-21 inhibitor group was significantly higher than that in the miR-21 NC group(P<0.01).The Western blot results showed that the expression of PDCD4 in the NC group(0.192±0.045)was significantly reduced compared to the miR-21 inhibitor group(0.683±0.091),and the expression level of the Bax protein in the NC group was 0.143±0.036,which was significantly reduced compared to the miR-21 inhibitor group(1.192±0.054),and the difference was also statistically significant(P<0.01).The expression of Bcl-2 protein in the NC group(0.864�

关 键 词：MIR-21 视网膜母细胞瘤 细胞增殖 凋亡 

分 类 号：R774[医药卫生—眼科]