Wnt5a抑制肺癌细胞系顺铂药物敏感性的作用研究

Wnt5a inhibit the sensitivity of cisplatin in the lung cancer cells

目的研究Wnt/PKC信号通过调控肺癌干细胞的特性对肺癌细胞系顺铂药物敏感性的影响。方法分别以化疗药物顺铂敏感和耐药的细胞系A549和A549/DDP为模型,用Wnt5a-CM激活或GF109203X抑制非经典Wnt信号,在顺铂刺激的情况下采用CCK8、细胞划痕实验对等实验分析细胞增殖、迁移等方面进行分析,用免疫印迹法对Wnt/PKC信号中相关蛋白及凋亡相关蛋白进行检测分析。结果 Wnt5a对A549和A549/DDP的增殖、迁移以及克隆能力都有促作用,免疫印迹法进一步分析发现Wnt5a通过激活Wnt/PKC信号通路来抑制肺癌细胞的部分凋亡,并且对该信号级联抑制可以降低肺癌干细胞的特性、促进细胞的凋亡,并通过caspase/AIF依赖途径来增加顺铂耐药细胞系A549/DDP对顺铂的再次敏感性。结论 Wnt5a通过激活Wnt/PKC信号通路来促进肺癌细胞的特性和顺铂耐药性。因此,阻断Wnt/PKC信号通路可能为化疗药物耐药NSCLC患者提供新的可供选择的治疗策略。

Objective To explore the inhibition of Wnt5 a sensitive to cisplatin in the lung cancer cells. Methods A549 cells were used as cisplatin sensitive model and A549/DDP cells as cisplatin resistant model,and treated by Wnt5 a conditional medium or protein kinase C( PKC) inhibitor GF109203 X,respectively. The properties of lung cancer stem cells and chemoresistance were accessed by cell proliferation,migration,and clonogenicity,the key proteins of Wnt/PKC and apoptosis signaling pathway were measured by Western blot. Results Compare to the control group,Wnt5 a promoted the proliferation,migration,and colony formation in A549 lung adenocarcinoma cells and cisplatin-resistant A549/DDP cells. GF109203 X strikingly increased the baseline apoptosis and resensitize the Wnt5 a-inhibited cell apoptosis. Interestingly,an inhibition of Wnt/PKC signaling pathway reduced the properties of lung cancer stemcells,promoted cell apoptosis,and resensitized cisplatin-resistant cells via a caspase/AIF-dependent pathway. Conclusion This results suggest that the Wnt5 a could promote lung cancer cell mobility and cisplatin-resistance through a Wnt/PKC signaling pathway and a blockage of this signaling may be an alternative therapeutic strategy for NSCLC patients with resistance to chemotherapies.