Atg3及自噬对非小细胞肺癌A549细胞迁移的影响

Effect of Atg3 and Autophagy on Migration of Non-small Cell Lung Cancer A549 Cells

目的探讨自噬相关基因Atg3和自噬对非小细胞肺癌A549细胞迁移的影响及相关机制。方法饥饿处理诱导非小细胞肺癌A549细胞自噬,Transwell法检测细胞迁移能力的变化,Western blot检测自噬相关基因Atg3的表达。转染Atg3 si RNA靶向沉默Atg3的表达,Transwell法检测细胞迁移能力的变化、流式细胞术检测细胞凋亡、Western blot检测细胞的自噬活性以及E-cadherin、β-catenin、N-cadherin、vimentin、Notch和snail的表达。结果饥饿处理可诱导非小细胞肺癌A549细胞自噬活性增强,并提高Atg3的表达,促进细胞迁移,而不影响细胞凋亡。沉默Atg3可抑制饥饿诱导的细胞自噬及迁移,而不影响细胞凋亡。此外沉默Atg3之后,E-cadherin和β-catenin的表达明显升高,而N-cadherin、vimentin、Notch和snail的表达明显降低。结论沉默Atg3可减弱自噬活性,抑制A549细胞的迁移;其机制可能与Notch/snail信号通路活性降低进而影响细胞的EMT进程有关。

Objective To investigate the effect of Atg3 and autophagy on the migration of non-small cell lung cancer cell line A549 and its possible mechanisms. Methods Starvation-induced autophagy was used in nonsmall cell lung cancer cell lines A549. Cell migration was calculated by Transwell assay and the expression of Atg3 was measured with Western blot. The Atg3 expression with Atg3 siRNA was down-regulated, and then cell migration was calculated by CCK-8 assay. Meanwhile, cell apoptosis was measured by flow cytometry, the autophagic activity and the expression of E-cadherin, 13-catenin, N-cadherin, vimentin, Notch and snail were detected by Western blot. Results Autophagic activities were effectively induced by starvation, with a stronger cell migration ability and higher Atg3 expression, while with no affection on cell apoptosis. The whole process of starvation-induced cell autophagy and migration could be suppressed by Atg3 knockdown; but Atg3 knockdown didn＇t affect cell apoptosis. Other than that, the knockdown of Atg3 was accompanied with higher E-cadherin and J3-catenin expression, and lower expression of N-cadherin, vimentin, Notch and snail. Conclusion The down-regulation of Atg3 could suppress cell autophagy and migration in non-small cell lung cancer. And its mechanism may be related to suppressing EMT progress via Notch/snail pathway.