lncRNA LINC02381通过miR-133a-3p/MGMT轴对脑胶质瘤细胞替莫唑胺抵抗的影响  

The effect of lncRNA LINC02381 on temozolomide resistance of glioma cells through the miR-133a-3p/MGMT axis

作　　者：陈静 董善武 陶双 罗超 陈玲 刘娜 付阳 CHEN Jing;DONG Shanwu;TAO Shuang;LUO Chao;CHEN Ling;LIU Na;FU Yang(Department of Pediatrics,Wuhan Fourth Hospital,Hubei Wuhan 430000,China)

机构地区：[1]武汉市第四医院儿科,湖北武汉430000

出　　处：《现代肿瘤医学》2023年第20期3751-3757,共7页Journal of Modern Oncology

基　　金：湖北省武汉市医学科研项目(编号:WX21C05)。

摘　　要：目的:探讨lncRNA LINC02381通过miR-133a-3p/MGMT轴对脑胶质瘤细胞替莫唑胺(TMZ)耐药性的影响。方法:体外培养人脑胶质瘤细胞株U251、耐TMZ细胞U251/TR,将U251/TR细胞分为U251/TR组、si-NC组、si-LINC02381组、mimics NC组、miR-133a-3p mimics组、miR-133a-3p mimics+pcDNA3.1组、miR-133a-3p mimics+pcDNA3.1 LINC02381组、miR-133a-3p mimics+pcDNA3.1 MGMT组。CCK-8法检测U251、U251/TR细胞对TMZ的耐药性;qRT-PCR检测LINC02381、miR-133a-3p、MGMT水平;流式细胞仪检测细胞凋亡;双荧光素酶报告实验验证LINC02381与miR-133a-3p、miR-133a-3p与MGMT的靶向关系;Western blot法检测Bax、Bcl-2、Cleaved-caspase 3、MGMT蛋白表达。结果:与U251细胞相比,U251/TR细胞中IC 50、LINC02381和MGMT mRNA、MGMT蛋白表达升高(P<0.05),miR-133a-3p表达降低(P<0.05);与U251/TR组、mimics NC组相比,miR-133a-3p mimics组Bcl-2表达降低(P<0.05),miR-133a-3p表达、细胞凋亡率、Bax和Cleaved-caspase 3表达升高(P<0.05);与U251/TR组、si-NC组相比,si-LINC02381组LINC02381、Bcl-2表达降低(P<0.05),细胞凋亡率、Bax和Cleaved-caspase 3表达升高(P<0.05);miR-133a-3p与LINC02381、MGMT均存在靶向关系;LINC02381、MGMT过表达均可逆转miR-133a-3p过表达对U251/TR细胞TMZ耐药性的影响。结论:lncRNA LINC02381沉默可能通过使miR-133a-3p表达升高,进而抑制MGMT表达,降低U251/TR细胞对TMZ的耐药性。Objective:To investigate the effect of lncRNA LINC02381 on the resistance of glioma cells to temozolomide(TMZ)through the miR-133a-3p/MGMT axis.Methods:Human glioma cell lines U251 and TMZ-resistant cells U251/TR were cultured in vitro,and U251/TR cells were divided into U251/TR group,si-NC group,si-LINC02381 group,mimics NC group,miR-133a-3p mimics group,miR-133a-3p mimics+pcDNA3.1 group,miR-133a-3p mimics+pcDNA3.1 LINC02381 group,and miR-133a-3p mimics+pcDNA3.1 MGMT group.CCK-8 method was performed to measure the resistance of U251 and U251/TR cells to TMZ.qRT-PCR was performed to measure the levels of LINC02381,miR-133a-3p,and MGMT.Flow cytometry was performed to measure cell apoptosis.Dual luciferase report experiment was performed to verify the targeting relationship between LINC02381 and miR-133a-3p,between miR-133a-3p and MGMT.Western blot method was performed to measure the protein expression of Bax,Bcl-2,Cleaved-caspase 3 and MGMT.Results:Compared with U251 cells,IC 50,the expression of LINC02381 and MGMT mRNA,MGMT protein expression in U251/TR cells increased(P<0.05),and the expression of miR-133a-3p decreased(P<0.05).Compared with U251/TR group and mimics NC group,the Bcl-2 expression in miR-133a-3p mimics group decreased(P<0.05),and the miR-133a-3p expression,cell apoptosis rate,the expression of Bax and Cleaved-caspase 3 increased(P<0.05).Compared with the U251/TR group and the si-NC group,the expression of LINC02381 and Bcl-2 in the si-LINC02381 group decreased(P<0.05),and the cell apoptosis rate,the expression of Bax and Cleaved-caspase 3 increased(P<0.05).miR-133a-3p had a targeting relationship with LINC02381 and MGMT.Both LINC02381 and MGMT overexpression could reverse the effect of miR-133a-3p overexpression on the TMZ resistance of U251/TR cells.Conclusion:The silencing of lncRNA LINC02381 may increase the expression of miR-133a-3p,thereby inhibiting the expression of MGMT and reducing the resistance of U251/TR cells to TMZ.

关 键 词：脑胶质瘤 lncRNA LINC02381 miR-133a-3p MGMT 

分 类 号：R739.41[医药卫生—肿瘤]