吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (05): 877-881.doi: 10.13481/j.1671-587x.20160507

• 基础研究 • 上一篇    下一篇

DNA-PKcs介导多药耐药恶性胶质瘤细胞化疗抗性及分子机制

惠双, 郭宏强   

  1. 河南省南阳市中心医院肿瘤内科, 河南 南阳 473009
  • 收稿日期:2016-04-19 出版日期:2016-09-28 发布日期:2016-09-29
  • 通讯作者: 惠双(Tel:0377-61660213,E-mail:nyhui1976@163.com) E-mail:nyhui1976@163.com
  • 作者简介:惠双(1976-),男,河南省南阳市人,副主任医师,医学硕士,主要从事肿瘤耐药性方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(81101797)

Chemoresistance of multi-drug resistance malignant glioma cells mediated by DNA-PKcs and its molecular mechanism

HUI Shuang, GUO Hongqiang   

  1. Department of Oncology, Nanyang Central Hospital, Henan Province, Nanyang 473009, China
  • Received:2016-04-19 Online:2016-09-28 Published:2016-09-29

摘要:

目的:观察DNA依赖的蛋白激酶的催化亚单位(DNA-PKcs)对多药耐药的恶性胶质瘤细胞化疗抗性的影响,探讨其介导化疗抗性的分子机制。方法:采用siRNA技术构建DNA-PKcs基因表达沉默的人胶质瘤U251细胞株;采用Western blotting法检测U251细胞(U251细胞)、多柔比星(ADM)耐药的U251细胞(U251/ADM细胞)和DNA-PKcs基因表达沉默的多柔比星耐药的U251细胞(U251/ADM/siDNA-PKcs细胞)中DNA-PKcs蛋白表达水平;采用CCK8法检测3组细胞增殖活性;采用Western blotting法检测3组细胞中多药耐药性1(MDR1)、报告基因质粒pNF-κB/p6、总Akt蛋白、pAkt/T308和pAKT/S473蛋白表达水平。结果:U251/ADM细胞DNA-PKcs蛋白表达水平明显高于U251细胞和U251/ADM/siDNA-PKcs细胞(P<0.01);ADM、紫杉醇(PTX)和吉西他滨(GEM)对U251/ADM/siDNA-PKcs细胞的半数抑制浓度(IC50)值较U251/ADM细胞明显降低(P<0.01);U251/ADM/SIDNA-PKcs细胞中pAKT/S473、pNF-κB/p65和MDR1蛋白表达水平均明显低于U251/ADM细胞(P<0.05),而两者总Akt和pAkt/T308蛋白表达水平比较差异无统计学意义(P>0.05)。结论:DNA-PKcs能明显增强多重耐药的恶性胶质瘤细胞化疗抗性,其作用机制与pAKT/S473和pNF-κB/p65表达上调,诱导MDR1表达有关。

关键词: DNA依赖的蛋白激酶, 胶质瘤, 化疗抗性, 多药耐药蛋白1

Abstract:

Objective: To obesrve the influence of DNA-PKcs in the chemoresistance of multi-drug resistance malignant glioma cells, and to explore its molecuIar mechanism in chemoresistance. Methods: siRNA was used to construct the DNA-PKcs knockdown human glioma U251 cell line; Western blotting method was used to detect the expressions of DNA-PKcs in U251 cells (U251 cells),doxorubicin (ADM) resistant U251 cells (U251/ADM cells), DNA-PKcs knockdown and ADM resistant U251 cells (U251/ADM/siDNA-PKcs cells).CCK8 method was used to detect the cell proliferation activity in three groups; Western blotting method was used to detect the expressions of MDR1, pNF-κB/p6, total Akt, pAkt/T308 and pAKT/S473 in the cells in three groups. Results: The expression level of DNA-PKcs in U251/ADM cells was significantly higher than those in U251 cells and U251/ADM/siDNA-PKcs cells (P<0.01). The IC50 values of doxorubicin (ADM), paclitaxel (PTX), gemcitabine (GEM) in U251/ADM/siDNA-PKcs cells were significantly lower than that in U251/ADM cells (P<0.05); the expression levels of pAKT/S473, pNF-κB/p65, and MDR1 in U251/ADM/siDNA-PKcs cells were significantly lower than those in U251/ADM cells (P<0.01), but the total Akt and pAkt/T308 had no significant differences between two groups (P>0.05). Conclusion: DNA-PKcs can significantly enhance the chemoresistance of multi-drug resistance malignant glioma cells, the underlying mechanism is related to up-regulation of pAKT/S473,pNF-κB/p65 and MDR1 expressions.

Key words: DNA-dependent kinase, glioma, chemoresistance, multi drug resistance protein 1

中图分类号: 

  • R739.4