基于凋亡和自噬途径的石榴皮多酚对人前列腺癌PC3细胞的抑制作用机制研究
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篇名: 基于凋亡和自噬途径的石榴皮多酚对人前列腺癌PC3细胞的抑制作用机制研究
TITLE: Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway
摘要: 目的:基于自噬和凋亡途径研究石榴皮多酚(PPP)对人前列腺癌PC3细胞增殖的抑制作用机制。方法:采用CCK-8法考察不同质量浓度PPP(25~300μg/mL)作用24、48、72h对PC3细胞活性的影响,筛选给药浓度和作用时间。将PC3细胞分为对照组(完全培养基)和PPP低、中、高质量浓度组,培养48h后,采用流式细胞术和AnnexinV-FITC/PI染色法分别检测细胞周期分布和凋亡情况,采用Westernblotting法检测细胞中凋亡相关蛋白Bax、Bcl-2和自噬相关蛋白LC3、Beclin-1、p62、Atg12、Atg16的表达水平。结果:选择48h为培养时间,PPP的半数抑制浓度为110μg/mL,选择50、100、200μg/mL为低、中、高质量浓度。与对照组比较,PPP低、中质量浓度组G0/G1期细胞百分比均显著降低而S期细胞百分比均显著升高,PPP高质量浓度组G0/G1期细胞百分比显著升高,PPP中、高质量浓度组G2/M期细胞百分比均显著降低(P<0.05或P<0.01);PPP各质量浓度组细胞凋亡率均显著升高(P<0.05或P<0.01)。与对照组比较,PPP各质量浓度组的抗凋亡蛋白Bcl-2和自噬相关蛋白p62表达水平均有不同程度的下降,促凋亡蛋白Bax表达水平和自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值和Beclin-1、Atg5、Atg12和Atg16的表达水平均有不同程度的上升,且PPP高质量浓度组上述指标以及低、中质量浓度组上述部分指标组间比较差异均有统计学意义(P<0.05或P<0.01)。结论:PPP能抑制人前列腺癌PC3细胞增殖,其机制可能与诱导细胞发生自噬从而促进细胞凋亡有关。
ABSTRACT: OBJECTIVE:To st udy the inhibitory mechanism of pomegranate peel polyphenols (PPP)on the proliferation of human prostate cancer PC 3 cells based on autophagy and apoptosis pathway. METHODS :CCK-8 assay was used to investigate the effects of PPP with different concentrations (25-300 μg/mL)on PC 3 cell activity after culturing for 24,48,72 h,so as to screen the drug concentration and treatment time. PC 3 cells were divided into control group (complete culture medium ),PPP low- ,medium- and high-concentration groups. After treated for 48 h,flow cytometry and Annexin V-FITC/PI staining were used to detect cell cycle distribution and apoptosis of PC 3 cells. Western blotting assay was used to detect the expression of apoptosis-related protein as Bax ,Bcl-2,as well as the expression of autophagy-related proteins as LC 3,Beclin-1,p62,Atg12 and Atg 16. RESULTS :The culturing time was chosen as 48 h. IC 50 of PPP was 110 μg/mL,and 50,100,200 μg/mL were chosen as low,medium,high concentrations of PPP. Compared with control group ,the percentage of PC 3 cells at phase G 0/G1 decreased significantly in PPP low- and medium-concentration groups while increased significantly at phase S ;that of PC 3 cells at phase G 0/G1 increased significantly in PPP high-concentration group ; while that of PC 3 cells at phase G 2/M decreased significantly in PPP medium- and high-concentration groups (P<0.05 or P<0.01). The apoptosis rate of PC 3 cells was increased significantly in PPP groups (P< 0.05 or P<0.01). Compared with control group ,protein expression of anti-apoptosis protein Bcl- 2 and autophagy-related promoting protein p 62 were decreased in PPP groups to different extents ,while protein expression of promoting-apoptosis protein Bax as wells as autophagy-related protein LC 3-Ⅱ/LC3-Ⅰ ration and protein expression of Beclin- 1,Atg5,Atg12 and Atg 16 were increased to different extents ;there was statistical significance in above indexes in PPP high-concentration group and some of above indexes in PPP low- and medium-concentration groups (P<0.05 or P<0.01). CONCLUSIONS :PPP can inhibit the proliferation of human prostate cancer PC 3 cells,mechanism of which may be related to inducing autophagy and promoting apoptosis.
期刊: 2020年第31卷第16期
作者: 冯甜,刘盟,程路峰,高晓黎,杨晓君
AUTHORS: FENG Tian,LIU Meng,CHENG Lufeng ,GAO Xiaoli,YANG Xiaojun
关键字: 石榴皮多酚;前列腺癌;PC3细胞;凋亡;自噬;机制
KEYWORDS: Pomegranate peel polyphenols ;Prostate cancer ;PC3 cell;Apoptosis;Autophagy;Mechanism
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