阿司匹林抑制HIF-1α表达和非小细胞肺癌上皮间质转化介导的细胞侵袭机制研究

Mechanism of Aspirin Mediating Cell Invasion via Inhibiting HIF-1αExpression and Epithelial-Mesenchymal Transition of Non-small Cell Lung Cancer

目的探讨阿司匹林通过抑制HIF-1α表达和非小细胞肺癌上皮间质转化介导的细胞侵袭的作用机制。方法选取非小细胞肺癌患者手术切除的癌组织和癌旁组织作为组织标本,Western blotting和实时荧光定量PCR检测E-cadherin和Vimentin的表达。培养非小细胞肺癌细胞株A549,分别加入0 mmol·L-1(对照组)、2.5 mmol·L-1(低剂量组)、5.0 mmol·L-1(中剂量组)和10.0 mmol·L-1(高剂量组)阿司匹林,Western blotting和实时荧光定量PCR检测细胞中E-cadherin、Vimentin和HIF-1α的表达。利用Transwell侵袭实验和划痕实验检测A549细胞的侵袭和迁移能力。结果与癌旁组织比较,癌组织中E-cadherin的mRNA及蛋白相对表达量明显升高,Vimentin的mRNA及蛋白相对表达量明显降低,差异均有统计学意义(P<0.05)。低、中、高剂量组HIF-1α、Vimentin的mRNA及蛋白相对表达量、侵袭细胞数及迁移距离均显著低于对照组,并随药物浓度升高而降低,组间差异均有统计学意义(P<0.05);低、中、高剂量组E-cadherin的mRNA及蛋白相对表达量显著高于对照组,并随药物浓度升高而升高,组间差异均有统计学意义(P<0.05)。结论细胞上皮间质转化与非小细胞肺癌病情进展相关,阿司匹林可通过下调HIF-1α表达和抑制细胞上皮间质转化介导肿瘤细胞的侵袭作用。

Objective To investigate the mechanism of aspirin mediating cell invasion by inhibiting HIF-1αexpression and epithelialmesenchymal transition of non-small cell lung cancer(NSCLC).Methods The cancer tissues and paracancerous tissues from NSCLC patients after operation were selected as tissue samples.The expression of E-cadherin and vimentin were detected by Western blotting and real-time fluorescence quantitative PCR.NSCLC cell line A549 were cultured,and were added with aspirin at doses of 0 mmol·L-1(control group),2.5 mmol·L-1(low dose group),5.0 mmol·L-1(medium dose group)and 10.0 mmol·L-1(high dose group).Western blotting and realtime fluorescence quantitative PCR were used to detect E-cadherin,vimentin and HIF-1αexpression in A549 cells.The invasion and migration of A549 cells were detected with Transwell invasion assay and cell scratch assay.Results Compared with the paracancerous tissues,the relative expression of E-cadherin protein and its mRNA in cancer tissues increased significantly,but the relative expression of vimentin protein and its mRNA decreased significantly(P<0.05).Compared with the control group,the relative protein and mRNA levels of HIF-1αand vimentin,the invasion cell number,and the migration distance were all decreased in low,medium and high dose groups,and they were decreased along with the increase of drug concentration(P<0.05).Whereas the relative protein and mRNA level of E-cadherin were higher in low,medium and high dose groups than in control group(P<0.05),and they were raised along with the increase of drug concentration(P<0.05).Conclusion Cell epithelial-mesenchymal transition was associated with non-small cell lung cancer.Aspirin could down-regulate the HIF-1αexpression and inhibit cell epithelial-mesenchymal transition to mediate cancer cell invasion.