木犀草素对人舌鳞状细胞癌Tca8113 细胞凋亡 及自噬的影响

Effects of luteolin on apoptosis and autophagy of human tongue squamous cell carcinoma Tca8113

摘要:

目的 探究木犀草素对人舌鳞状细胞癌Tca8113 细胞体外凋亡和自噬作用的影响。方法 采 用MTT 法观察12.5、25.0、50.0 及100.0μmol/L 的木犀草素作用24 h 对细胞体外活性的影响；运用 Hoechst33342 染色法和流式细胞术观察体外细胞凋亡情况；Western blotting 检测Cleaved Caspase-3 蛋白、 B 细胞淋巴瘤-2（Bcl-2）、B 细胞淋巴瘤-2 相关X 蛋白（Bax）和Beclin1 蛋白、P62 蛋白及LC3B 蛋白。 结果 MTT 法实验结果显示，木犀草素抑制Tca8113 细胞增殖，IC50 为47μmol/L。Hoechst33342 染色结果 显示，木犀草素使Tca8113 细胞形态发生改变，与时间呈正相关（P <0.05）。流式细胞术结果表明，47μmol/L 的木犀草素作用Tca8113 细胞12、24 h 后，凋亡率高于空白对照组（P <0.05）。Western bloting 检测结果表明， 木犀草素抑制Bcl-2 以及增加Bax 表达，导致Cleaved Caspase-3 表达增加，促进Tca8113 细胞凋亡（P <0.05）； 同时还升高Beclin1、LC3B、P62 蛋白水平（P <0.05）。结论 木犀草素影响人舌鳞状细胞癌Tca8113 细胞凋 亡和自噬的发生。

Abstract:

Objective To investigate the effect of luteolin (Luteolin, Lut) on apoptosis and autophagy in human tongue squamous cell carcinoma Tca8113 cells in vitro. Methods MTT assay was used to observe the effects of (12.5, 25, 50, 100 μmol/L) Lut on cell activity in vitro after 24 hours. Heochst33342 and flow synectometry were used to observed cell apoptosis. Western blotting revealed changes in the protein expression of Cleaved Caspase-3, b-lymphocytoma-2 (Bcl-2), b-lymphocytoma-2-related X protein (Bax), and Beclin1, P62 and LC3B proteins. Results MTT assay results showed that Lut significantly inhibited the proliferation of Tca8113 cells with IC50 of 47 mol/L. Hoechst33342 staining showed that Lut could change the morphology of Tca8113 cells, which was positively correlated with time. Annexin V/PI double staining results showed that the apoptosis rate of Tca8113 cells treated with 47 mol/L Lut for 12h and 24h was higher than that of the control group (P < 0.05). Western Bloting showed that Lut inhibited Bcl-2 and increased Bax expression, leading to increasimg Cleaved Caspase-3 expression and promoting Tca8113 cell apoptosis (P < 0.05). Beclin1, LC3B and P62 protein levels were also increased (P < 0.05). Conclusion Apoptosis and autophagy of tongue cancer cell line Tca8113 may be caused by Luteolin.