雷公藤红素抑制人喉癌Hep-2细胞增殖并诱导其凋亡

Proliferation-inhibiting and apoptosis-inducing effects of tripterine on human laryngeal carcinoma Hep-2 cells

目的研究雷公藤红素对人喉癌Hep-2细胞增殖及凋亡的影响,并初步探讨其可能的作用机制。方法以不同浓度、不同时间的雷公藤红素处理体外培养的人喉癌Hep-2细胞,采用锥虫蓝(台盼蓝)染色、CCK-8和结晶紫染色法检测其对细胞增殖的影响;DAPI荧光染色法和流式细胞术检测细胞凋亡情况;Western blot检测EZH2、H3K27me3的蛋白表达变化。结果雷公藤红素能够抑制喉癌Hep-2细胞的增殖且呈时间与剂量依赖性,48h后其IC50为(3.19±0.83)μmol/L;与对照组相比,5μmol/L的雷公藤红素作用24h后Hep-2凋亡率显著增加(t=-11.32,P<0.05),EZH2、H3K27me3蛋白表达水平下降。结论雷公藤红素能显著抑制人喉癌Hep-2细胞的增殖并诱导其凋亡,可能是通过下调EZH2及下游靶基因H3K27me3从而发挥作用。

Objective To explore the effects and mechanism of tripterine on the proliferation and apoptosis of human laryngeal carcinoma Hep-2 cells. Methods Human laryngeal carcinoma Hep-2 cells were treated with tripterine of different concentrations and in different days. The effects on cell proliferation were measured by trypan blue staining, CCK-8 method and crystal violet staining. The effects on cell apoptosis were observed by the fluorescence dye DAPI and flow cytometry. The protein expressions of EZH2 and H3K27me3 were detected by Western blot. Results Tripterine inhibited the proliferation of human laryngeal carcinoma Hep-2 cells in time- and dose-dependent manners; IC50 was （3.19 ＋ 0.83）/~mol/L at 48 hours. Tripterine of 5 /zmol/L could promote karyopyknosis and fragmentation of Hep-2 cell nucleus at 24 hours. The apoptosis rate was increased compared with that in control group （t =- 11. 32, P〈0.05）. The protein expressions of EZH2 and H3K27me3 were down- regulated. Oonclusion Tripterine can significantly inhibit the proliferation and induce the apoptosis of human laryngeal carcinoma Hep-2 cells in vitro, which may be related to down-regulation of EZH2 and H3K27me3 expres- sions.