小檗碱通过调控PI3K/Akt通路抑制喉癌细胞生长和转移

Berberine inhibits growth and metastasis of laryngeal carcinoma cells by regulating PI3K/Akt pathway

目的探讨小檗碱对喉癌Hep2细胞增殖、凋亡和侵袭的影响以及相关作用机制。方法培养Hep2细胞分为空白对照组和5、10、20μmol·L-1小檗碱组,CCK-8检测Hep2的增殖活性,流式细胞术检测细胞凋亡情况,划痕实验检测细胞迁移情况,Transwell法检测细胞侵袭能力,Western blot法检测凋亡蛋白酶激活因子1(Apaf1)、剪切后半胱天冬酶(cl-caspase)-9、cl-caspase-3蛋白、血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-2、MMP-9、磷脂酰肌醇3激酶(PI3K)、磷酸化蛋白激酶B(p-Akt)和p-p65蛋白表达水平。结果与空白对照组相比,5、10、20μmol·L-1小檗碱组细胞增殖活性显著降低(P <0.01);凋亡细胞百分比显著增加(P <0.01),Apaf1、cl-caspase-9和cl-caspase-3蛋白水平显著上调(P <0.01);划痕愈合率显著降低(P <0.01),侵袭细胞数目显著减少(P <0.01);细胞中VEGF、MMP-2、MMP-9、PI3K、p-Akt、p-p65蛋白水平显著降低(P <0.01),且均呈浓度依赖性。结论小檗碱通过调控PI3K/Akt通路抑制喉癌细胞增殖、侵袭和迁移,诱导细胞凋亡。

AIM To investigate the effects of berberine on proliferation,apoptosis and invasion of laryngeal cancer Hep2 cells and its related mechanisms. METHODS Cultured Hep2 cells were divided into blank control group and 5, 10 and 20 μmol·L-1 berberine groups.The proliferative activity and apoptosis of Hep2 cells were detected by CCK-8 and flow cytometry. The cell migration and invasion ability were detected by Scratch test and Transwell assay. The protein expression of apoptotic protease activator 1(Apaf1), cleaved caspase-9(cl-caspase-9), cl-caspase-3, vascular endothelial growth factor(VEGF), matrix metalloproteinases(MMP)-2, MMP-9, phosphatidylinositol 3 kinase(PI3K), phosphorylated protein kinase B(p-Akt) and p-p65 were detected by Western blot. RESULTS Compared with the blank control group,the cell proliferation activity of berberine 5, 10 and 20 μmol·L-1 groups were significantly decreased(P < 0.01). The percentage of apoptotic cells were increased(P < 0.01), and the protein levels of Apaf1, cl-caspase-9 and cl-caspase-3 were increased(P < 0.01). The scratch healing rate was significantly decreased(P < 0.01), and the number of invasive cells was decreased(P < 0.01). The protein levels of VEGF, MMP-2, MMP-9, PI3K, p-Akt and p-p65 in cells were significantly decreased(P < 0.01), and all indexes were in concentration-dependent manner. CONCLUSION Berberine can inhibit the proliferation, invasion and migration of laryngeal cancer cells and induce apoptosis by regulating PI3K/Akt pathway.