miR-145-5p通过TCTP调节胶质瘤细胞对DMC-BH的耐药性  

MiR-145-5p regulates the drug resistance of glioma cells to DMC-BH through TCTP

作　　者：程超 孙关 石磊 CHENG Chao;SUN Guan;SHI Lei(Department of Critical Care Medicine, Kunshan Hospital Affiliated to Jiangsu University/Gusu College of Nanjing Medical University, Kunshan 215300, China;Department of Neurosurgery, Yancheng First People's Hospital, Yancheng 224008, China;Department of Neurosurgery, Kunshan Hospital Affiliated to Jiangsu University/ Gusu College of Nanjing Medical University, Kunshan 215300,China)

机构地区：[1]昆山市第一人民医院/南京医科大学姑苏学院急诊医学科,江苏昆山215300 [2]盐城市第一人民医院神经外科,江苏盐城224008 [3]昆山市第一人民医院/南京医科大学姑苏学院神经外科,江苏昆山215300

出　　处：《中国肿瘤外科杂志》2022年第3期278-284,共7页Chinese Journal of Surgical Oncology

基　　金：国家自然科学基金(81370062)。

摘　　要：目的 探讨miR-145-5p对胶质母细胞瘤耐DMC-BH细胞株的作用及其调节机制。方法 建立对DMC-BH耐药的胶质母细胞瘤细胞株U87/DMC-BH和U251/DMC-BH。实时荧光定量PCR检测DMC-BH耐药细胞株中miR-14-5p表达水平。采用CCK-8和流式细胞分别检测miR-145-5p过表达对U87/DMC-BH和U251/DMC-BH细胞活性和凋亡的影响。利用Western blot检测MRP1和MDR1等蛋白变化。结果 miR-145-5p在耐药株U87/DMC-BH和U251/DMC-BH细胞表达低于非耐药细胞株U87和U251,分别为后者的32.3%和22.1%。过表达miR-145-5p降低了耐药株U87和U251细胞的增殖活性,72 h时间点细胞存活率分别为对照组46.5%和38.4%,并诱导其凋亡增加。Western blot检测提示耐药相关基因MRP1和MDR1表达也显著减少。TCTP蛋白在U87/DMC-BH和U251/DMC-BH中过表达;但转染agomiR-145-5p后,TCTP表达显著下降。TCTP siRNA可显著抑制U87/DMC-BH和U251/DMC-BH中TCTP表达,并抑制其细胞增殖、促进凋亡。而共转染TCTP过表达载体和agomiR-145-5p后逆转了agomiR-145-5p对细胞的增殖抑制和凋亡诱导,并上调了MRP1和MDR1表达。结论 miR-145-5p通过靶向TCTP增强了耐药胶质瘤细胞对DMC-BH的敏感性。Objective To investigate the effect and regulatory mechanism of miR-145-5p on the resistance to DMC-BH in glioblastoma cell line.Methods Glioblastoma cell lines U87/DMC-BH and U251/DMC-BH resistant to DMC-BH were established.The expression level of miR-14-5p in DMC-BH resistant cell line was detected by real-time fluorescence quantitative PCR.CCK-8 and flow cytometry were used to detect the effects of miR-145-5p overexpression on the activity and apoptosis of U87/DMC-BH and U251/DMC-BH cells,respectively.The changes of protein expression of MRP1 and MDR1 were detected by Western blot.Results The expression of miR-145-5p in drug-resistant U87/DMC-BH and U251/DMC-BH cells was lower than that in non-resistant cells U87 and U251 accounting for 32.3%and 22.1%of the latter,respectively.Overexpression of miR-145-5p reduced the proliferation activity of drug-resistant U87 and U251 cells.And the cell viability of U87 and U251 cells at 72 h was 46.5%and 38.4%of the control group,respectively,which induced an increase in apoptosis.Western blot detection showed that the expression of drug resistance-related genes MRP1 and MDR1 were also significantly reduced.TCTP was overexpressed in U87/DMC-BH and U251/DMC-BH,which was significantly decreased after transfection with agomiR-145-5p.The expression of TCTP in U87/DMC-BH and U251/DMC-BH was inhibited by TCTP siRNA significantly which inhibited cell proliferation and promoted apoptosis as well.However,co-transfection of TCTP overexpression vector and agomiR-145-5p reversed the inhibition of cell proliferation and induction of apoptosis by agomiR-145-5p,and up-regulated the expressions of MRP1 and MDR1.Conclusions miR-145-5p enhances the sensitivity of drug-resistant glioma cells to DMC-BH by targeting TCTP.

关 键 词：miR-145-5p TCTP DMC-BH 胶质瘤 耐药 

分 类 号：R739.4[医药卫生—肿瘤]