肺积宁方影响Lewis肺癌细胞增殖和自噬的作用机制研究

Feijining Decoction on Proliferation and Autophagy in Lewis Cells

目的:研究肺积宁方对Lewis肺癌细胞增殖和自噬的影响及相关信号通路的探讨。方法:选用不同浓度的肺积宁方(FJND)提取物作用于体外培养的Lewis肺癌细胞,MTT法检测FJND对Lewis细胞增殖的抑制作用;通过GFP-LC3自噬定位观察细胞自噬情况;蛋白免疫印迹法检测不同浓度FJND作用Lewis细胞后,Beclin-1蛋白、LC3B-Ⅰ、LC3B-Ⅱ、Atg5蛋白表达;和m TOR,AKT,p-AKT相关蛋白表达情况。结果:FJND在3.12 mg/m L抑制Lewis细胞增殖作用最强,Beclin-1蛋白、Atg5蛋白表达及LC3B-Ⅱ/LC3B-Ⅰ比值明显高于对照组;对m TOR,AKT,p-AKT蛋白抑制作用强于对照组。结论:肺积宁方抑制细胞增殖和诱导自噬,可能通过抑制PI3K/Akt/m TOR信号通路发挥作用。

Objective： To observe Feijining Decoction（ FJND） inhibiting proliferation and inducing autophagy in Lewis cells and the possible related signaling pathway. Methods： Lewis cell cultures were incubated with various concentrations of FJND for different duration. The MTT assay was used to assess the compounds ability to inhibit cellular proliferation.Autophagy. GFP- LC3 positioning was used to identify autophagic vacuoles. The protein levels of Beclin- 1,LC3B- Ⅰ,LC3B- Ⅱ,Atg5 and m TOR,AKT,p- AKT were determined by Western blotting so as to evaluate the molecular pathways of autophagy and the related signaling pathway. Results： At concentrations of 3. 12 mg / m L,FJND significantly inhibited proliferation and increased autophagy. And Lewis cells treated with FJND increased expressions of Beclin- 1,LC3B-Ⅱ / LC3B- Ⅰ and Atg5 and the expression levels of m TOR,Akt and p Akt were significantly lower in the Lewis cells than those in the untreated control cells. Conclusion： FJND inhibits proliferation and induces autophagy in Lewis cells via regulating the PI3 k / Akt / m TOR pathway.