基于PI3K/AKT信号通路探讨双氢青蒿素对人三阴性乳腺癌MDA-MB-231细胞增殖、凋亡的影响

徐嘉若; 陈佳靓; 姚广涛

doi:10.16306/j.1008-861x.2021.06.008

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基于PI3K/AKT信号通路探讨双氢青蒿素对人三阴性乳腺癌MDA-MB-231细胞增殖、凋亡的影响

实验研究 | 更新时间：2022-09-09

- 基于PI3K/AKT信号通路探讨双氢青蒿素对人三阴性乳腺癌MDA-MB-231细胞增殖、凋亡的影响
- Effects of dihydroartemisinin on proliferation and apoptosis of human triple-negative breast cancer MDA-MB-231 cells based on PI3K/AKT signaling pathway
- 上海中医药大学学报 2021年35卷第6期页码：49-57
- 作者机构：
  
  1.上海中医药大学（上海　201203）
- 作者简介：
  
  徐嘉若，女，硕士，主要从事中药新药及药理学研究
  姚广涛，副研究员，硕士生导师；E-mail：yaoguangtao1969@126.com
- 基金信息：
  
  国家科技重大专项项目(2019ZX09201004-002);上海市教委预算内项目(18LK019)
- DOI：10.16306/j.1008-861x.2021.06.008
  中图分类号：
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徐嘉若, 陈佳靓, 姚广涛. 基于PI3K/AKT信号通路探讨双氢青蒿素对人三阴性乳腺癌MDA-MB-231细胞增殖、凋亡的影响[J]. 上海中医药大学学报, 2021,35(6):49-57.

XU Jiaruo, CHEN Jialiang, YAO Guangtao. Effects of dihydroartemisinin on proliferation and apoptosis of human triple-negative breast cancer MDA-MB-231 cells based on PI3K/AKT signaling pathway[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(6):49-57.
徐嘉若, 陈佳靓, 姚广涛. 基于PI3K/AKT信号通路探讨双氢青蒿素对人三阴性乳腺癌MDA-MB-231细胞增殖、凋亡的影响[J]. 上海中医药大学学报, 2021,35(6):49-57. DOI： 10.16306/j.1008-861x.2021.06.008.

XU Jiaruo, CHEN Jialiang, YAO Guangtao. Effects of dihydroartemisinin on proliferation and apoptosis of human triple-negative breast cancer MDA-MB-231 cells based on PI3K/AKT signaling pathway[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(6):49-57. DOI： 10.16306/j.1008-861x.2021.06.008.

摘要

目的：,2,基于PI3K/AKT信号通路探讨双氢青蒿素（DHA）对人三阴性乳腺癌MDA-MB-231细胞增殖及凋亡的影响。,方法：,2,①不同浓度（25、50、100 μmol/L）DHA干预不同时间（12、24、48、72 h）后，MTT法检测细胞增殖。②不同浓度（0、30、60、90 μmol/L）DHA干预48 h后，流式细胞仪检测细胞凋亡和细胞周期，Hoechst 33258染色观察细胞凋亡。③划痕实验观察60 μmol/L DHA干预0、6、12、24 h后对细胞迁移能力的影响。④60 μmol/L DHA干预12 h后，PCR检测细胞内凋亡相关因子,Caspase-3,、,Caspase-9,、,Bcl-2,、,Bax,的mRNA表达水平。60 μmol/L DHA干预48 h后，Western blot检测细胞内Caspase-3、Caspase-9、Bcl-2、Bax以及PI3K、p-PI3K、AKT、p-AKT的蛋白表达水平。,结果：,2,①DHA能够抑制MDA-MB-231细胞增殖，且抑制作用随药物浓度增加而增强。②DHA可以促进MDA-MB-231细胞凋亡，且随着药物浓度增加，促凋亡作用增强。③30 μmol/L DHA作用可使细胞周期阻滞在G0/G1期；60、90 μmol/L DHA作用后细胞周期阻滞在G2/M期。④60 μmol/L DHA干预12、24 h后，细胞迁移率显著降低（,P,<,0.01）。⑤60 μmol/L DHA干预后，,Caspase-3,、,Caspase-9, mRNA表达水平以及,Bax,/,Bcl-2, mRNA表达比值显著上调（,P,<,0.01），cleaved Caspase-9/Caspase-9、Bax/Bcl-2蛋白表达比值显著升高（,P,<,0.05）；同时PI3K蛋白表达水平显著升高（,P,<,0.05），而p-PI3K、p-AKT的蛋白表达水平显著降低（,P,<,0.01）。,结论：,2,DHA可能通过抑制PI3K/AKT信号通路，抑制MDA-MB-231细胞的增殖和迁移，并诱导细胞凋亡。

Abstract

Objective:,2,To investigate the effects of dihydroartemisinin(DHA) on the proliferation and apoptosis of human triple-negative breast cancer MDA-MB-231 cells based on PI3K/AKT signaling pathway.,Methods:,2,①After treatment with DHA at different concentrations(25, 50, 100 μmol/L) for different times(12, 24, 48, 72 h), the cell proliferation was detected by MTT assay.②After treatment with DHA at different concentrations(0, 30, 60, 90 μmol/L) for 48 hours, the cell apoptosis and cell cycle were detected by flow cytometry, and the cell apoptosis was observed by Hoechst 33258 staining.③The effect of 60 μmol/L DHA on the cell migration was observed by scratch test after treatment for 0, 6, 12 and 24 hours respectively.④After treatment with DHA at 60 μmol/L for 12 hours, the mRNA expression levels of apoptosis related factors ,Caspase-3,Caspase-9,Bcl-2, and ,Bax, were detected by PCR.After treatment with DHA at 60 μmol/L for 48 hours, the protein expression levels of Caspase-3, Caspase-9, Bcl-2, Bax, PI3K, p-PI3K, AKT and p-AKT were detected by Western blot.,Results:,2,①DHA could inhibit the proliferation of MDA-MB-231 cells, and the inhibitory effect was increased with the increase of drug concentration.②DHA could promote the apoptosis of MDA-MB-231 cells, and this effect was increased with the increase of drug concentration.③DHA at 30 μmol/L could block the cell cycle in G0/G1 phase, and DHA at 60 and 90 μmol/L could block the cell cycle in G2/M phase.④After treatment with DHA at 60 μmol/L for 12 and 24 hours, the cell migration rate was decreased significantly(,P,<,0.01) .⑤After treatment with DHA at 60 μmol/L, the expression levels of ,Caspase-3, and ,Caspase-9, mRNA and the expression ratio of ,Bax/Bcl-2, mRNA were significantly up-regulated(,P,<,0.01), and the expression ratios of cleaved Caspase-9/Caspase-9 and Bax/Bcl-2 protein were significantly increased(,P,<,0.05) ; meanwhile, the protein expression level of PI3K was increased significantly(,P,<,0.05), and the protein expression levels of p-PI3K and p-AKT were decreased significantly(,P,<,0.01) .,Conclusion:,2,DHA may inhibit the proliferation and migration of MDA-MB-231 cells and induce the cell apoptosis by inhibiting PI3K/AKT signaling pathway.

关键词

双氢青蒿素三阴性乳腺癌细胞增殖细胞凋亡PI3K/AKT信号通路MDA-MB-231细胞

Keywords

dihydroartemisinintriple-negative breast cancercell proliferationcell apoptosisPI3K/AKT signaling pathwayMDA-MB-231 cell

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