免疫抑制剂FK506对肺癌细胞增殖和迁移的作用研究

摘要

Background and objective FK506, also named tacrolimus, a new macrolide immunosuppressive agent, has been shown to possess anti-proliferation activities in some cancer cells. The aim of this study was to investigate the effect of FK506 on the cell proliferation and migration of lung cancer cell lines and its mechanism. Methods A549 and H1299 cell lines were cultured in vitro. The effect of FK506 on cell viability and DNA synthesis ability of A549 and H1299 were measured by CCK-8 assay and EDU-labeling assay, respectively. Flow cytometry assay was used to detect the cell cycle. The in vitro mi-gration of lung cancer cells was detected by Boyden chamber assay and wound-healing assay after the treatment of FK506. The expression of p27, RB1, CDK4, CDK6 and MMP9 were detected using Western blot. Results FK506 inhibited cell growth and induced cell cycle arrest in G0/G1 phase in A549 and H1299 cells in a dose- and time-dependent manner. Compared to the control groups, the migration of A549 and H1299 cells treated with FK506 were decreased obviously. Moreover, FK506 in-creased the expression of P27 and RB1, and reduced the expression of CDK4, CDK6 and MMP9. Conclusion FK506 inhibit the cell growth and migration of lung cancer cells in vitro. The inhibitive effects may be associated with the up-regulation of p27 expression and inhibition CDK4, CDK6 and MMP9 expression.%背景与目的 FK506(他克莫司,tacrolimus)是一种大环内酯类的新型免疫抑制剂.研究报道FK506对多种肿瘤细胞具有增殖抑制作用.本研究旨在观察FK506对肺癌细胞增殖和迁移的作用,并探讨其可能的机制.方法 体外培养A549和H1299细胞,采用CCK-8法测定FK506对A549和H1299细胞增殖的作用;EDU标记法检测DNA合成;流式细胞术检测细胞的周期分布情况;Transwell和细胞划痕实验检测细胞的体外迁移能力;Western blot技术检测P27、RB1、CDK4、CDK6和MMP9蛋白的表达.结果 FK506可抑制A549和H1299细胞的增殖、诱导细胞周期G0期/G1期阻滞;与对照组比较,FK506处理后A549和H1299细胞迁移能力明显降低,且呈剂量依赖性.此外,与对照组相比,FK506处理组中P27和RB1表达上调,而CDK4、CDK6和MMP9表达显著下降.结论 FK506对人肺癌A549和H1299细胞的增殖和迁移能力有明显的抑制作用,其机制可能与上调p27表达、抑制CDK4、CDK6和MMP-9表达有关.