双氢青蒿素联合吉非替尼抑制肺腺癌细胞周期和迁移能力的体外研究

Combination of dihydroartemisinin and gefitinib inhibits cell cycle and migration of lung adenocarcinoma cells in vitro

目的探究双氢青蒿素(dihydroartemisinin,DHA)联合吉非替尼(gefitinib,GEF)对肺腺癌细胞系的细胞周期和迁移能力的影响。方法 10μM DHA和(或) 50μM GEF处理肺腺癌细胞系A549和SPC-A-1,四唑盐(methyl thiazolyl tetrazolim,MTT)比色法检测细胞增殖,流式细胞术检测细胞周期,划痕实验检测细胞迁移,蛋白质印记法(western blot,WB)检测单独或联合应用DHA和GEF对细胞周期和迁移相关蛋白表达的影响。结果 MTT实验结果显示联合应用DHA和GEF(Com组)后,A549和SPC-A-1的增殖能力显著低于单药组和对照(NC)组,且具有时间依赖性(P <0. 05);碘化丙啶(propidium iodide,PI)单染流式细胞术检测细胞阻滞于G0/G1期; WB显示与NC组和单药组相比,Com组细胞周期蛋白依赖性蛋白激酶4(cyclin dependent kinase 4,CDK4)和细胞周期素D1(Cyclin D1)表达显著下降;划痕实验中Com组细胞融合率显著低于NC组和单药组,并伴随基质金属蛋白酶(matrix metalloproteinase,MMP) 2和MMP9表达下降,差异均具有统计学意义(P <0. 05)。结论 DHA联合GEF可以显著抑制肺腺癌细胞的增殖和迁移,并阻滞细胞周期。

Objective To explore the combined effect of dihydroartemisinin( DHA) and gefitinib( GEF) in lung adenocarcinoma cell lines. Methods Lung adenocarcinoma cell lines A549 and SPC-A-1 were treated with 10μM DHA and( or) 50μM GEF.MTT assay was used to detect cell proliferation,while flow cytometry was adopted to determine cell cycle. Cell migration was observed with cell scratch tests. The effects of DHA and GEF alone or in combination on the expressions of proteins related to cycle and migration were detected by Western blot. Results MTT results showed that DHA combined with GEF( Com group) inhibited the proliferation of A549 and SPC-A-1 cell lines more significantly than in control and single drug groups and in a time-dependent manner( P <0. 05). Flow cytometry with PI staining showed that cell cycles were blocked at G0/G1 phase. Western blot showed that the expressions of CDK4 and Cyclin D1 in the Com group decreased more significantly than in control and single drug groups. Cell fusion rate of Com group was also significantly lower than that of control and single drug groups in cell scratch test and the expressions of MMP2 and MMP9 were decreased. The difference was statistically significant( P < 0. 05). Conclusions DHA combined with GEF can significantly inhibit the proliferation and migration of lung adenocarcinoma cells and block cell cycle.