冬虫夏草繁育品对恶性黑色素瘤B16细胞增殖及迁移能力的影响

Effects of cultivated Cordyceps sinensis on proliferation and migration of B16 melanoma cells

目的探讨冬虫夏草Cordyceps sinensis繁育品对恶性黑色素瘤B16细胞增殖及迁移能力的影响。方法通过MTT法、平板克隆实验检测冬虫夏草繁育品对B16细胞增殖能力的影响;采用划痕实验、transwell小室实验检测其对B16细胞迁移能力的影响;通过黏附实验检测其对B16细胞黏附能力的影响;采用流式细胞仪检测其对B16细胞周期分布的影响;采用Western blotting法检测其对B16细胞中MMP-2、MMP-9、Bax、Bcl-2、Cyclin D1、CDK2、CDK4、P21及p-Akt蛋白表达的影响。结果冬虫夏草繁育品能够将B16细胞阻滞在G1/S期,从而显著抑制B16细胞的增殖,并显著降低B16细胞的迁移能力。同时可导致B16细胞MMP-9、MMP-2、Bcl-2、Cyclin D1、CDK2、CDK4、p-Akt蛋白表达减少和Bax、P21蛋白表达增加。结论冬虫夏草繁育品能显著抑制B16细胞的增殖及迁移能力,其作用机制可能与调控细胞周期相关蛋白、MMPs家族蛋白及p-Akt蛋白的表达有关。

Objective To investigate the effects of cultivated Cordyceps sinensis on inhibiting the proliferation and migration of B16 melanoma cells. Methods MTT assay and clone formation assay were used to examine the inhibitory effect ofcukivated C. sinensis on the proliferation of B16 cells; Scratching test and transwell assay were used to detect its effect on migration; Cell adhesion assay was used to detect its effect on adhesion. Flow cytometry was used to detect its effect on cell cycle arrest; Western blotting was used to detect its effect on the expression of the proteins of MMP-2, MMP-9, Bax, Bcl-2, CyclinD 1, CDK2, CDK4, P21, and p-Akt. Results The results showed that cultivated C. sinensis can significantly inhibit the proliferation and clone formation of B16 ceils via arresting cells at G-/S phase in a dose-dependent manner. Moreover, cell migration was also substantially inhibited in a dose-dependent manner. Western blotting analysis showed that cultivated C. sinensis obviously increased the levels of Bax and P21, and meanwhile, decreased the expression of Bcl-2, MMP-2, MMP-9, CDK2, CDK4, CyclinD1, and p-Akt. Conclusion The inhibitory effects of cultivated C. Sinensis on the proliferation and migration orb 16 cells probably associated with the expression of related regulating proteins, MMPs family proteins and p-Akt protein in cell cycle.