Twist通过促进MRP3表达诱导肺癌细胞A549耐药的研究

Twist induces drug resistance in human lung cell line A549 by up-regulating MRP3

目的研究Twist增强A549细胞顺铂耐药性的分子机制。方法 CCK8法检测Twist对A549细胞对于顺铂的敏感性;RT-PCR和Western blotting检测Twist对A549细胞中耐药基因表达的影响。免疫组化检测肺癌组织中Twist和MRP3表达相关性。结果在顺铂(32,64,128μmol/L)作用下,过表达Twist和对照组相比,生存率分别为(0.79±0.039、0.63±0.048、0.46±0.039 vs.0.53±0.039、0.41±0.043、0.27±0.063),差异具有统计学意义。发现Twist可以促进MRP3表达,敲除MRP3表达后逆转了Twist诱导的耐药性。临床数据显示,Twist和MRP3在肺癌组织中表达具有相关性(P<0.05)。结论 Twist通过上调MRP3促进肺癌细胞A549对顺铂的耐药性。

Objective To investigate the molecular mechanism of Twist induced-drug resistance in human lung cell line A549 cells.Methods The sensibility of A549 cells over-expressed Twist to cisplatin was measured by CCK8 assay. The expressions of Twist,MRP1,MRP2 and MRP3 were detected by RT-PCR. Western blotting was used to detect the expressions of Twist and MRP3 in A549 cells over-expressed Twist.The correlation between Twist and MRP3 expressions in 30 cases of lung cancer tissues was detected with immunohistochemistry. Results The viability of A549 cells over-expressed Twist was significant better than the control group under cispla-tin（32,64,128 μmol/L）treatment（0.79 ± 0.039,0.63 ± 0.048,0.46 ± 0.039 vs.0.53 ± 0.039,0.41 ± 0.043, 0.27 ± 0.063,respectively）. After knockdown of MRP3 expression,it reversed drug resistance induced by Twist. Clinically,the expressions of Twist and MRP3 in lung cancer tissues were significantly relevant（P 〈 0.05）. Conclusions Twist induces the drug resistance of human lung cell line A549 to cisplatin via up-regulating the expression of MRP3.