EPHA10促进肺腺癌H1975奥希替尼耐药细胞增殖、耐药和迁移的实验研究

Experimental study of EPHA10 promoting proliferation,drug resistance and migration of lung adenocarcinoma H1975 ocitinib-resistant cells

目的研究EPHA10在H1975奥希替尼耐药细胞增殖、耐药和迁移中的作用及机制。方法通过Western Blot检测EPHA10在表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)敏感和耐药细胞株中的表达情况,选取EPHA10高表达细胞株作为研究对象。利用小RNA干扰技术降低高表达株中EPHA10表达,分别采用CCK-8,Transwell实验检测对照组和干扰组中细胞的增殖、耐药及迁移能力,再利用Western Blot检测干扰EPHA10后对细胞增殖和迁移关键蛋白的影响。结果EPHA10在H1975奥希替尼耐药细胞中表达最高;在该细胞中干扰EPHA10后,细胞的增殖、耐药及迁移能力下降,且细胞增殖相关的关键蛋白p-ERK、PCNA、Cyclin D1以及迁移相关蛋白ZEB1、Slug、MMP9、Vimentin表达均下调。结论敲低EPHA10抑制了肺腺癌H1975奥希替尼耐药细胞的增殖、耐药和迁移,其作用机制可能与抑制细胞中ERK通路激活和细胞转移有关。

Objective To investigate the role and mechanism of EPHA10 in the proliferation,resistance,and migration of osimertinib-resistant H1975 cells.Methods The expression of EPHA10 in epidermal growth factor receptor tyrosine kinase inhibitors(EGFR-TKIs)sensitive and drug-resistant cell lines was detected by Western Blot,and EPHA10 high-expression cell lines were selected as the study object.Small RNA interference technology was used to reduce the expression of EPHA10 in high-expression strains.CCK-8 and Transwell experiments were used to detect the proliferation,drug resistance,and migration ability of cells in the control group and the interference group,respectively.Western Blot was used to detect the effects of interference with EPHA10 on key proteins of cell proliferation and migration.Results The expression of EPHA10 was the highest in H1975 ohitinib resistant cells.After interfering with EPHA10 in this cell,the proliferation,drug resistance and migration ability of the cells were decreased,and the expressions of key proteins related to cell proliferation p-ERK,PCNA,Cyclin D1,and migration-related proteins ZEB1,Slug,MMP9,and Vimentin were down-regulated.Conclusion EPHA10 knockdown can inhibit the proliferation,drug resistance,and migration of osimertinib-resistant lung adenocarcinoma H1975 cells,which may be related to the inhibition of ERK pathway and metastasis.