Src激酶抑制剂对人肺癌顺铂耐药细胞A549/DDP多药耐药性的影响及机制

Role and mechanism of Src tyrosine kinase inhibitor on multi-drug resistance of human cis-platinum-resistant lung cancer cell line A549/DDP

目的研究Src激酶抑制剂PP2对人肺癌顺铂耐药细胞A549/DDP多药耐药性的影响及机制。方法以5和10μmol.L-1Src激酶抑制剂PP2作用A549/DDP细胞24h后,应用Western blot考察肿瘤细胞Src磷酸化表达的变化,MTT法检测细胞的药物敏感性,流式细胞仪考察细胞P-gp表达的变化,Western blot及Real-time PCR考察肿瘤细胞MDR1蛋白及mRNA表达的变化。结果 Src激酶抑制剂PP2可下调A549/DDP细胞Src磷酸化表达,5和10μmol.L-1 Src激酶抑制剂PP2作用后,顺铂对A549/DDP细胞的药物敏感性分别提高了1.37和2.47倍,细胞P-gp表达分别为对照组的65.2%和46.4%,MDR1在蛋白水平表达显著降低,MDR1在mRNA水平的表达分别为对照组的50.24%(P<0.05)和37.6%(P<0.05)。结论 Src激酶抑制剂PP2可逆转A549/DDP细胞多药耐药性,其机制可能与降低细胞MDR1表达水平有关。

Objective To investigate the effect and mechanism of the Src kinase inhibitor PP2 on the multi-drug resistance of human cis-platinum-resistant lung cancer cell line A549/DDP.Methods After treatment of A549/DDP cells with 5 and 10 μmol·L-1 Src kinase inhibitor PP2 for 24 h,Western blot was used to investigate the change of the tumor cell Src phosphorylation,MTS assay was used to examine the anti-tumor drug sensitivity change,and flow cytometry was used to investigate the P-gp expression alteration,Western blot and real-time PCR was used to investigate the change of the tumor cell MDR1 protein and mRNA expression.Results The Src kinase inhibitor PP2 could down-regulate the Src phosphorylation in A549/DDP cells,after treatment with 5 and 10 μmol·L-1 PP2.The cis-platinum sensitivity increased 1.37-fold and 2.47-fold for A549/DDP cells respectively.And the cellular P-glycoprotein（P-gp） expression was 65.2%,and 46.4% of the control respectively.The protein expression of MDR1 was significantly decreased,and the mRNA expression of MDR1 was 50.24% and 37.6% of the control respectively.Conclusion Src kinase inhibitor PP2 could reverse the multi-drug resistance in A549/DDP cells,and the mechanism may involve the down-regulation of the cell MDR1 expression.