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目的:探究不同剂量柚皮素(naringenin)对人下咽癌FADU细胞的作用及其相关机制。方法:以不同剂量(0、100、200、400、800μg/m L)柚皮素对FADU细胞进行处理,倒置显微镜观察细胞形态学变化;cck-8法检测药物作用后不同时间(24、48、72 h)FADU细胞增殖的变化;流式细胞仪检测柚皮素作用48 h细胞凋亡的变化;采用Transwell趋化小室体外侵袭实验测定48 h后细胞的侵袭能力的变化;蛋白免疫印迹法检测48 h后细胞内PI3K/AKT信号通路凋亡相关蛋白表达的影响。结果:cck-8法检测发现柚皮素抑制FADU细胞增殖作用明显,呈浓度和时间依赖性;流式细胞结果发现200,400,800μg/m L柚皮素作用48 h后,FADU细胞的凋亡率显著增高,差异有统计学意义(P<0.05)。Transwell体外侵袭实验发现,随着柚皮素浓度增加,FADU细胞穿透聚碳酸酯滤膜的能力逐渐减弱,柚皮素有效的抑制FADU细胞在体外的侵袭能力,呈浓度依赖性。同时,蛋白免疫印迹法凋亡相关蛋白检测结果表明,FADU细胞内BCL2、P-AKT和P-PI3K蛋白表达均受到明显抑制。结论:柚皮素能有效抑制人下咽癌FADU细胞的增殖、体外侵袭能力,诱导细胞凋亡;其机制与下调PI3K/AKT信号通路凋亡相关蛋白表达有关。
Objective: To investigate the effects of naringenin with different dosages on human hypopharyngeal carcinoma FADU cells and its related mechanisms. Methods: FADU cells were treated with different concentrations of naringenin (0, 100, 200, 400 and 800 μg / mL), morphological changes of the cells were observed under inverted microscope, cck- , 72 h) FADU cell proliferation; Flow cytometry was used to detect the changes of apoptosis induced by naringenin at 48 h; The invasion ability of cells was measured by Transwell chemotaxis chamber in vitro 48 h; Western blotting The effect of PI3K / AKT signaling pathway on apoptosis related protein expression 48 h after the test. Results: Cynanisin inhibited the proliferation of FADU cells in a dose-dependent and time-dependent manner by flow cytometry. The apoptosis rate of FADU cells treated with 200, 400 and 800 μg / mL of naringenin for 48 h was significantly higher Increased, the difference was statistically significant (P <0.05). Transwell in vitro invasion assay showed that with the increase of naringenin concentration, the ability of FADU cells to penetrate the polycarbonate membrane gradually weakened, and naringenin effectively inhibited the invasion ability of FADU cells in vitro in a concentration-dependent manner. At the same time, Western blotting results showed that the expression of BCL2, P-AKT and P-PI3K in FADU cells were significantly inhibited. Conclusion: Naringenin can effectively inhibit the proliferation of human hypopharyngeal carcinoma FADU cells in vitro, and induce the apoptosis of FADU cells in vitro. The mechanism is related to the down-regulation of the expression of apoptosis-related proteins in PI3K / AKT signaling pathway.