苯丁酸钠联合吉非替尼对肺腺癌 A549细胞增殖、迁移的影响及机制

摘要

Objective To study the effects of sodium phenylbutyrate combined with gefitinib on proliferation and mi -gration of lung adenocarcinoma A 549 cells and to investigate the mechanism .Methods The lung adenocarcinoma A 549 cells were cultured and divided into the blank control group ( treated with DMEM medium without FBS ) , gefitinib group (treated with 1 mmol/L gefitinib), sodium phenylbutyrate group (treated with 5 μmol/L sodium phenylbutyrate ) and the combined treatment group ( treated with 1 mmol/L gefitinib and 5 mol/L sodium phenylbutyrate ) .After culture for 24 h, the OD450 was detected by MTT method and the cell cycle distribution was detected by flow cytometry .The expression of ap-optosis-related genes survivin , BRAF, Notch1, MMP-1, MMP-2, MMP-3 and MMP-9 was detected by RT-PCR.The cell migration was assayed by scratch test .Results Compared with the blank control group , the OD450 , cell proportion in S phase, cell mobility and the expression of survivin , BRAF, Notch1, MMP-1, MMP-2, MMP-3 and MMP-9 mRNA were decreased, and the cell proportion in G0/G1 phase was increased in the combined treatment group , sodium phenylbutyrate group and gefitinib group (all P<0.05).Compared with the sodium phenylbutyrate group and gefitinib group , the OD450, cell proportion in S phase , cell mobility and the expression of survivin , BRAF, Notch1, MMP-1, MMP-2, MMP-3 and MMP-9 mRNA were decreased , and the cell proportion in G 0/G1 phase was increased in the combined treatment group ( all P<0.05).Conclusion The sodium phenylbutyrate and gefitinib can both inhibit the proliferation and migration process of lung adenocarcinoma , which may be related with the decreased expression of survivin , BRAF, Notch1, MMP-1, MMP-2, MMP-3 and MMP9 mRNA.The combined application of the two drugs has synergistic effect .%目的：研究苯丁酸钠联合吉非替尼对肺腺癌A549细胞增殖、迁移的影响，并探讨其机制。方法将体外培养的肺腺癌A549细胞分为空白对照组、吉非替尼组、苯丁酸钠组、联合处理组，各组均加入不含FBS的DMEM培养基，另外吉非替尼组加1 mmol／L吉非替尼，苯丁酸钠组加5μmol／L苯丁酸钠，联合处理组加1 mmol／L吉非替尼和5μmol／L苯丁酸钠。培养24 h后，用MTT法检测各组细胞OD450值，用流式细胞术检测各组细胞周期分布，用RT-PCR法检测凋亡相关基因survivin、BRAF、Notch1、MMP1、MMP2、MMP3、MMP9 mRNA，行划痕实验检测细胞迁移能力。结果与空白对照组相比，联合处理组、苯丁酸钠组、吉非替尼组OD450值、S期细胞比例、细胞迁移率及凋亡相关分子survivin、BRAF、Notch1、MMP1、MMP2、MMP3、MMP9 mRNA降低，G0／G1期细胞比例升高，P均＜0．05。与苯丁酸钠组、吉非替尼组相比，联合处理组OD450值、S期细胞比例、细胞迁移率及凋亡相关分子survivin、BRAF、Notch1、MMP1、MMP2、MMP3、MMP9 mRNA降低，G0／G1期细胞比例升高，P均＜0．05。结论苯丁酸钠和吉非替尼均能抑制肺腺癌细胞的增殖和迁移过程，其机制可能与降低细胞survivin、BRAF、Notch1、MMP1、MMP2、MMP3、MMP9 mRNA表达有关，两种药物联合应用具有协同作用。