重组人Rab17通过调节STAT3/HIF-α/VEGF信号通路影响非小细胞肺癌增殖和侵袭

Effect of recombinant human Rab17 on proliferation and invasion of non-small cell lung cancer by regulating STAT3/HIF-1α/VEGF signaling pathway

目的探讨RAS癌基因家族中重组人Rab17对非小细胞肺癌(Non-small cell lung cancer,NSCLC)细胞增殖、迁移和侵袭的影响及可能的作用机制。方法qRT-PCR和Western blot实验检测NSCLC细胞中Rab17 mRNA和蛋白质表达水平;转染实验将Rab17质粒和对照质粒转染至A549细胞中,采用嘌呤霉素筛选出稳定表达的细胞株;MTT和软琼脂集落形成实验检测Rab17对A549细胞的增殖的影响;Rab17对A549细胞的迁移和侵袭的影响采用迁移和侵袭实验检测;Western blot实验检测Rab17对STST3、HIF-1α和VEGF蛋白表达的影响。结果三种NSCLC细胞系A549、H460和HCC827中Rab17 mRNA的相对表达水平和蛋白表达水平均低于正常人支气管上皮细胞Bease-2B;Rab17质粒成功转染至A549细胞中,并筛选出稳定表达的细胞株;Rab17可显著抑制A549细胞的增殖、迁移和侵袭;Rab17可抑制p-STAT3、HIF-α和VEGF蛋白的表达。结论Rab17在NSCLC细胞中表达水平降低,其可能是通过抑制STAT3/HIF-α/VEGF信号通路的传导抑制A549细胞的增殖、迁移和侵袭。

Objective To evaluate the effect of recombinant human Rab17 in the RAS oncogene family on the proliferation,migration and invasion of non-small cell lung cancer(NSCLC)cells and its possible mechanism.Methods qRT-PCR and Western blot were used to detect the expression of Rab17 mRNA and protein in non-small cell lung cancer cells.It used transfection experiments to transfect Rab17 plasmid and control plasmid into A549 cells,and puromycin to select stable expression cell lines.MTT and soft agar colony formation experiments were used to detect the proliferation of Rab17 on A549 cells.Migration and invasion experiments were used to detect the effect of Rab17 on the migration and invasion ability of A549 cells.Western blot experiment was used to detect the effect of Rab17 on the expression of STST3,HIF-1αand VEGF proteins.Results The relative expression levels of Rab17 mRNA in three kinds of NSCLC cells A549,H460 and HCC827 were lower than those of normal human bronchial epithelial cells Bease-2B.The expression intensity of Rab17 protein in the three kinds of NSCLC cells was weaker than that of Bease-2B cells.The Rab17 plasmid was successfully transfected into A549 cells,and stable-expressing cell lines were selected.Rab17 could significantly inhibit the proliferation,migration,and invasion of A549 cells.Rab17 could inhibit the expression of p-STAT3,HIF-α,and VEGF proteins.Conclusion Rab17 expression is reduced in NSCLC cells,which may be through inhibiting STAT3/HIF-α/VEGF signaling pathway to inhibit the proliferation,migration and invasion of A549 cells.