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.
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.)
出处
《现代肿瘤医学》
CAS
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)。
