Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway

FENG Tian; LIU Meng; CHENG Lufeng; GAO Xiaoli; YANG Xiaojun

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Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway

更新时间：2022-06-21

- Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway
- China Pharmacy Vol. 31, Issue 16, (2020)
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- Published：2020，
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FENG Tian, LIU Meng, CHENG Lufeng, et al. Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway. [J]. China Pharmacy 31(16).(2020)
DOI：

FENG Tian, LIU Meng, CHENG Lufeng, et al. Study on the Inhibitory Mechanism of Pomegranate Peel Polyphenols on Human Prostate Cancer PC 3 Cells Ba- sed on Apoptosis and Autophagy Pathway. [J]. China Pharmacy 31(16).(2020) DOI：

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.

关键词

石榴皮多酚前列腺癌PC3细胞凋亡自噬机制

Keywords

Pomegranate peel polyphenolsProstate cancerPC3 cellApoptosisAutophagyMechanism

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