MTDH干扰促进胶质瘤替莫唑胺耐药株的药物敏感性和细胞凋亡  

MTDH interference promotes drug sensitivity and cell apoptosis in glioma resistant to temozolomide

作　　者：孟亮 王跃飞 陶亮 樊文 涂勤 祝源 余小祥 戴小琴 MENG Liang;WANG Yuefei;TAO Liang;FAN Wen;TU Qin;ZHU Yuan;YU Xiaoxiang;DAI Xiaoqin(Department of Neurosurgery;2120 Treatment Center,Wuhan Third Hospital,Hubei Wuhan 430061,China.)

机构地区：[1]武汉市第三医院神经外科 [2]武汉市第三医院120救治中心,湖北武汉430061

出　　处：《现代肿瘤医学》2024年第14期2530-2534,共5页Journal of Modern Oncology

基　　金：湖北省武汉市科技局2022年度知识创新专项基础研究项目(编号:2022020801010549)。

摘　　要：目的:探讨异黏蛋白(metadherin, MTDH)干扰是否通过提高胶质瘤替莫唑胺(temozolomide, TMZ)耐药株的药物敏感性促进细胞凋亡。方法:基于培养的U251人神经胶质瘤细胞和U251MG/TMZ人星形胶质瘤耐替莫唑胺细胞,通过qRT-PCR检测细胞中MTDH、miR-1471和miR-153-3p的表达。根据人MTDH的序列合成siRNA/NC,并转染至U251MG/TMZ细胞中。通过CCK8检测细胞的增殖能力,流式细胞术检测细胞的凋亡,Western blot法检测耐药相关基因的表达。采用不同浓度的替莫唑胺(2.5、5、10、20、40、80、160μmol/L)作用于转染si-MTDH/NC的U251MG/TMZ细胞,48 h后通过CCK8法检测细胞增殖。结果:U251MG/TMZ细胞中具有高表达的MTDH和低表达的miR-1471、miR-153-3p。与Control组比较,U251MG/TMZ组细胞的增殖能力显著增加,凋亡率显著降低,MTDH、ABCG2、MGMT和Pgp的mRNA表达显著增加,MTDH、ABCB1、MGMT、ABCG2和p-AKT1的蛋白表达显著增加。与NC组比较,si-MTDH组细胞的增殖能力显著降低,凋亡率显著增加,MTDH、ABCG2、MGMT和Pgp的mRNA表达显著降低,MTDH、ABCB1、MGMT、ABCG2和p-AKT1的蛋白表达显著降低。结论:MTDH干扰通过提高胶质瘤替莫唑胺耐药株的药物敏感性促进细胞凋亡,其机制可能与AKT1通路有关。Objective:To investigate whether metadherin(MTDH)interference can promote cell apoptosis by improving the drug sensitivity of glioma temozolomide(TMZ)-resistant strains.Methods:Based on the cultured U251 human glioma cells and U251MG/TMZ human astroglioma cells resistant to temozolomide,the expressions of MTDH,miR-1471 and miR-153-3p in the cells were detected by qRT-PCR.siRNA/NC was synthesized according to the sequence of human MTDH and transfected into U251MG/TMZ cells.Cell proliferation ability was detected by CCK8.Cell apoptosis was detected by flow cytometry.The expression of drug resistance-related genes was detected by Western blot.U251MG/TMZ cells transfected with si-MTDH/NC were treated with different concentrations of temozolomide(2.5,5,10,20,40,80,160μmol/L),and cell proliferation was detected by CCK8 method after 48 h.Results:U251MG/TMZ cells had high expression of MTDH,low expression of miR-1471 and miR-153-3p.Compared with the Control group,the proliferation ability of cells in the U251MG/TMZ group was significantly increased,the apoptosis rate was significantly reduced,the mRNA expressions of MTDH,ABCG2,MGMT and Pgp were significantly increased,and the protein expressions of MTDH,ABCB1,MGMT,ABCG2 and p-AKT1 were significantly increased.Compared with the NC group,the proliferation ability of cells in the si-MTDH group was significantly decreased,the apoptosis rate was significantly increased,the mRNA expressions of MTDH,ABCG2,MGMT and Pgp were significantly decreased,and the protein expressions of MTDH,ABCB1,MGMT,ABCG2 and p-AKT1 were significantly decreased.Conclusion:MTDH interference promotes cell apoptosis by increasing the drug sensitivity of glioma resistant to temozolomide,and the mechanism may be related to the AKT1 pathway.

关 键 词：胶质瘤 MTDH 凋亡 药物敏感性 AKT1通路 

分 类 号：R739.41[医药卫生—肿瘤]