吉非替尼联合二甲双胍对EGFR-TKIs原发性耐药非小细胞肺癌H1975细胞的抑制作用及其机制研究

发布时间：2018-09-16 21:02

【摘要】：目的探讨吉非替尼联合二甲双胍对肺癌H1975细胞的抑制增效作用及其机制。方法采用MTT、克隆形成实验观察吉非替尼与二甲双胍单用及联用对原发性耐药非小细胞肺癌H1975细胞增殖活性的影响;Matrigel包被的Transwell小室检测细胞侵袭能力;划痕愈合实验检测细胞迁移能力;TUNEL检测细胞凋亡率;Western blot、免疫荧光法检测凋亡和上皮细胞-间充质转化(epithelial to mesenchymal transition,EMT)相关蛋白的变化。结果吉非替尼联合二甲双胍组对H1975细胞增殖、侵袭及迁移抑制效应优于吉非替尼组,协同增效作用明显(P0.05);吉非替尼和二甲双胍单用时较对照组的细胞凋亡率增加,当两者联合作用时细胞凋亡率增加更显著(P0.05),且明显下调抗凋亡相关蛋白(Bcl-xl、Mcl-1)和上调促凋亡蛋白Bax;两药联用明显逆转H1975细胞EMT的发生;间质细胞相关蛋白(Vimentin、N-cadherin、Slug、Snail)表达水平降低,上皮细胞相关蛋白E-cadherin表达水平升高。结论吉非替尼联合二甲双胍可显著抑制EGFR-TKIs原发性耐药非小细胞肺癌H1975细胞的增殖、侵袭和迁移,促使肿瘤细胞凋亡;其机制可能通过激活线粒体凋亡途径及逆转EMT的发生来发挥抗肿瘤作用。
[Abstract]:Objective to investigate the inhibitory and synergistic effects of gefitinib combined with metformin on lung cancer cell line H1975 and its mechanism. Methods the effect of gifitinib and metformin on the proliferative activity of H1975 cells in primary drug-resistant non-small cell lung cancer was observed by MTT, clone forming assay. The invasion ability of H1975 cells was detected by Transwell chamber coated by Matrigel. Cell migration ability was detected by scratch healing assay and apoptosis rate was detected by Tunel. Western blot, immunofluorescence assay was used to detect apoptosis and the changes of epithelial cell-mesenchymal transformation (epithelial to mesenchymal transition,EMT) related proteins. Results the inhibitory effect of gefitinib combined with metformin on the proliferation, invasion and migration of H1975 cells was better than that of gefitinib group, and the synergistic effect was significant (P0.05), and the apoptotic rate of gifitinib and metformin alone was higher than that of the control group. The apoptosis rate of H1975 cells increased more significantly (P0.05) when combined with the two drugs, and the expression of interstitial cell associated protein (Vimentin,N-cadherin,Slug,Snail) decreased, and the combination of anti-apoptosis-related protein (Bcl-xl,Mcl-1) and up-regulated apoptosis-promoting protein (Bax;) significantly reversed the occurrence of EMT in H1975 cells. The expression of epithelial associated protein E-cadherin was increased. Conclusion Gefitinib combined with metformin can significantly inhibit the proliferation, invasion and migration of H1975 cells in primary drug-resistant non-small cell lung cancer (EGFR-TKIs) and promote the apoptosis of tumor cells. The mechanism may play an antitumor role by activating mitochondrial apoptosis pathway and reversing the occurrence of EMT.
【作者单位】：第三军医大学大坪医院呼吸内科;
【分类号】：R734.2

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