甲基莲心碱通过p62抑制肺腺癌A549细胞侵袭、转移的机制★

The mechanisms of neferine in inhibiting the invasion and metastasis of lung adenocarcinoma A549 cells via p62★

摘要:

目的 阐明甲基莲心碱（Nef）抑制肺腺癌A549细胞侵袭和转移的可能机制，为今后临床应用Nef防治肺腺癌转移提供理论基础。方法 常规培养肺腺癌A549细胞，加入不同浓度的Nef进行干预，CCK-8检测细胞活力；将A549细胞分为三组：空白组不做任何处理，TGF-β组采用TGF-β1干预，TGF-β+Nef组采用TGF-β1和Nef干预，显微镜下观察细胞形态；Transwell和Wound healing法检测各组细胞的侵袭和迁移能力； PCR、Western blotting检测各组细胞上皮间质转化（EMT）相关基因和蛋白的表达，电镜下验证细胞自噬，p62基因沉默进一步明确作用机制。结果 Nef可明显抑制TGF-β1诱导的A549细胞EMT，并可能通过抑制Twist1自噬降解抑制A549细胞的侵袭和迁移。结论 Nef可抑制肺腺癌A549细胞的侵袭和转移，其机制可能与促进p62介导的Twist1选择性自噬降解有关。

Abstract:

Objective To elucidate the possible mechanism of neferine (Nef) in inhibiting the invasion and metastasis of lung adenocarcinoma A549 cells and to provide theoretical basis for the clinical application of Nef in the prevention and treatment of lung adenocarcinoma metastasis.Methods The lung adenocarcinoma A549 cells were cultured routinely and treated with different concentrations of Nef. The proliferation of A549 cells was detected by CCK-8. A549 cells were divided into three groups: blank group without any treatment, TGF-β group with TGF-β1 intervention, TGF-β+Nef group with TGF-β and Nef intervention. The morphology of A549 cells was observed under microscopes, and the invasion and migration ability of A549 cells in each group were detected by Transwell and scratch assay. The expressions of epithelial-mesenchymal transition (EMT) -related proteins and genes were detected by Western blotting and PCR. Electron microscopy confirmed the existence of autophagy. p62 gene knockout further clarified the mechanism.Results Nef significantly inhibited the EMT of A549 cells induced by TGF-β1. Nef may inhibit the invasion and metastasis of A549 cells by suppressing the autophagy degradation of Twist1.Conclusion Nef can inhibit the invasion and metastasis of lung adenocarcinoma and its mechanism may be related to the promotion of p62-mediated selective autophagy degradation of Twist1.