目的:探索不同浓度雷公藤红素(celastrol)对下咽癌FADU细胞增殖抑制及促进其凋亡的机制。方法:用不同浓度(0μmol/L,1μmol/L,3μmol/L,5μmol/L)雷公藤红素作用于FADU细胞,倒置显微镜下观察雷公藤红素干预后细胞形态变化情况;CCK-8法检测不同时间段(24、48、72h)药物干预后FADU细胞的增殖变化;流式细胞仪检测雷公藤红素干预24h后细胞凋亡率变化;Western—Blot检测不同药物浓度作用24h后FADU细胞P13K信号通路中与凋亡相关的关键蛋白的表达变化情况。结果:与对照组相比,在倒置显微镜下FADU细胞形态明显改变,细胞数量明显减少,细胞边界模糊不清,漂浮细胞增多,而随着用药浓度的增加细胞的增殖能力被显著抑制,P13K、AKT、P—mTOR、Bcl-2蛋白表达明显减少,并且肿瘤细胞的凋亡率明显增高。结论:雷公藤红素对FADU细胞具有显著的增殖抑制效果,同时能够促进细胞凋亡,其原理主要与通过下调P13K信号通路凋亡相关蛋白的表达有关。
Objective: To explore the Effects ofcelastrol on proliferation and apoptosis in FADU cells and its action mechanisms. Methods: The changes of FADU cells were observed after treatde at concentrations of 0 μmol/L, 1 μmol/L, 3 μmol/L, 5 μmol/L for 24 h, using the inverted microscope. Using the CCK-8 method, the effects of celastrol on the viability of FADU cells was measured after treated in different dose and different time periods (24, 48, 72 h). Using the flow cytometry, the effects of celastrol on the apoptosis of FADU cells was measured treated in different dose. Using the western-blot method, the key protein expression of PI3Ksignal pathway related to apoptosis was measured. Results: Compared with the control group, the alterations of the cells were obvious, FADU cells decreased significantly, the boundary of the cells is ambiguity, floating cells increased, and the viability, the expression of PI3K, AKT, P-mTOR, Bcl-2 decreased with the increase of dose and the apoptosis rate increases with the dose increased significantly. Conclusions: The celastrol has inhibitory effect on the viability of FADU cells and induced apoptosis of cells. Its mechanism could be related to the reduction of protein expression in PI3K signal pathway.