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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、72 h)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24 h后细胞凋亡率变化;Western-Blot检测不同药物浓度作用24 h后FADU细胞PI3K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,PI3K、AKT、P-m TOR、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PI3K信号通路凋亡相关蛋白的表达有关。
Objective: To explore the mechanism of different concentrations of celastrol on proliferation inhibition and apoptosis of hypopharyngeal cancer FADU cells. Methods: FADU cells were treated with different doses of tripterine (0μmol / L, 1μmol / L, 3μmol / L, 5μmol / L). The changes of cell morphology were observed under inverted microscopy. The proliferation of FADU cells was detected after 24 h, 48 h and 72 h of drug intervention. The changes of apoptotic rate of FADU cells were detected by flow cytometry after 24h intervention. After FADU cells PI3K signaling pathway and apoptosis-related key protein expression changes. Results: Compared with the control group, the morphological changes of FADU cells under inverted microscope were significantly changed, the number of cells was significantly reduced, the cell borders were blurred and the number of floating cells increased. With the increase of drug concentration, the proliferation of FADU cells was significantly inhibited. The expression of PI3K, AKT, Pm TOR, Bcl-2 protein expression was significantly reduced, and the rate of apoptosis of tumor cells was significantly increased. It is concluded that tripterine inhibits the proliferation of FADU cells and promotes the apoptosis of FADU cells. The principle is mainly related to the down-regulation of the expression of apoptosis-related proteins in PI3K signaling pathway.