甘 霖,熊志峰,杨化超,王鸿林,王 婷,吕 钢.槲皮素对乳腺癌MCF-7和MDA-MB-435细胞的凋亡作用及机制研究[J].,2021,(19):3638-3644 |
槲皮素对乳腺癌MCF-7和MDA-MB-435细胞的凋亡作用及机制研究 |
Apoptosis Effect and Mechanism Study of Quercetin on the Breast Cancer MCF-7 and MDA-MB-435 Cells |
投稿时间:2021-03-28 修订日期:2021-04-23 |
DOI:10.13241/j.cnki.pmb.2021.19.008 |
中文关键词: 乳腺癌 槲皮素 增殖 凋亡 Fas FasL Bcl-2 Bax |
英文关键词: Breast cancer Quercetin Proliferation Apoptosis Fas FasL Bcl- 2 Bax |
基金项目:重庆市2018年科研机构绩效激励引导专项项目(cstc2018jxj1130063) |
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中文摘要: |
摘要 目的:探讨槲皮素对乳腺癌MCF-7和MDA-MB-435细胞的凋亡作用,并探讨其作用机制。方法:采用活细胞计数法(CCK-8)测定槲皮素对乳腺癌MCF-7和MDA-MB-435细胞增殖的作用,分别采用细胞划痕实验和Transwell实验测定槲皮素对乳腺癌MCF-7和MDA-MB-435细胞迁移和侵袭的影响,采用流式细胞术测定槲皮素对乳腺癌MCF-7和MDA-MB-435细胞凋亡的作用,采用实时荧光定量聚合酶链反应(qRT-PCR)和免疫印迹法(West-blotting)测定槲皮素对乳腺癌MCF-7和MDA-MB-435细胞Fas、FasL、Bcl-2和Bax mRNA和蛋白表达的影响。结果:槲皮素(50~200 μmol/L)作用乳腺癌MCF-7和MDA-MB-435细胞 24 h、48 h和72 h对其增殖具有显著的抑制作用,并且呈浓度依赖性(P<0.05);细胞划痕实验中槲皮素50 μmol/L和100 μmol/L可使乳腺癌MCF-7和MDA-MB-435细胞划痕宽度较对照组显著增加(P<0.05);Transwell试验中槲皮素50 μmol/L和100 μmol/L可使乳腺癌MCF-7和MDA-MB-435穿膜细胞较对照组显著降低(P<0.05);槲皮素50 μmol/L和100 μmol/L可使乳腺癌MCF-7和MDA-MB-435细胞凋亡率较对照组显著升高(P<0.05);槲皮素50 μmol/L和100 μmol/L可使乳腺癌MCF-7和MDA-MB-435细胞中Bcl-2 mRNA表达较对照组显著降低(P<0.05),Fas、FasL和Bax mRNA表达较对照组显著升高(P<0.05);槲皮素50 μmol/L和100 μmol/L可使乳腺癌MCF-7和MDA-MB-435细胞中Bcl-2 蛋白表达较对照组显著降低(P<0.05),Fas、FasL和Bax 蛋白表达较对照组显著升高(P<0.05)。结论:槲皮素可促进乳腺癌细胞的凋亡,可能与其通过作用Fas/FasL凋亡信号通路而激活外源性凋亡途径,通过作用Bcl-2凋亡信号通路而激活内源性凋亡途径有关。 |
英文摘要: |
ABSTRACT Objective: To explore the apoptosis effect of quercetin on the breast cancer MCF-7 and MDA-MB-435 cells, and to explore its mechanism. Methods: The cell counting kit-8 (CCK 8) method was used to determine the effect of quercetin on proliferation of breast cancer MCF-7 and MDA-MB-435 cells, the cell scratch test and the Transwell test were respectively used to determine the effect of quercetin on migration and invasion of breast cancer MCF-7 and MDA-MB-435 cells. Flow cytometry was used to determine the effect of quercetin on apoptosis in breast cancer MCF-7 and MDA-MB-435 cells. The real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and West-blotting (West-blotting) were used to determine the effect of quercetin on mRNA and protein expression of Fas, FasL, Bcl-2 and Bax in breast cancer MCF-7 and MDA-MB-435 cells. Results: Quercetin (50~200 μmol/L) significantly inhibited the proliferation of breast cancer MCF-7 and MDA-MB-435 cells for 24 h, 48 h and 72 h, and it was concentration dependent manner (P<0.05). The cell scratch width of breast cancer MCF-7 and MDA-MB-435 cells by quercetin 50 μmol/L and 100 μmol/L were significantly increased in cell scratch test compared with those in control group (P<0.05). In Transwell test, the penetrating cells of breast cancer MCF-7 and MDA-MB-435 by quercetin 50 μmol/L and 100 μmol/L were significantly reduced compared with those in the control group(P<0.05). Compared with those in the control group, the cell apoptosis rate of breast cancer MCF-7 and MDA-MB-435 by quercetin 50 μmol/L and 100 μmol/L were significantly increased(P<0.05). Compared with those in the control group, the expression of Bcl-2 mRNA of breast cancer MCF-7 and MDA-MB-435 cells by quercetin 50 μmol/L and 100 μmol/L were significantly decreased (P<0.05), while the expressions of Fas, FasL and Bax mRNA were significantly increased compared with those in the control group (P<0.05). The protein expression of Bcl-2 of breast cancer MCF-7 and MDA-MB-435 cells by quercetin 50 μmol/L and 100 μmol/L were significantly reduced compared with those in the control group (P<0.05), while the protein expression of Fas, FasL and Bax were significantly increased compared with those in the control group (P<0.05). Conclusion: Quercetin can promote the apoptosis of breast cancer cells, which may be related to the activation of exogenous apoptosis pathway by acting on the Fas/FasL apoptosis pathway, and the activation of endogenous apoptosis pathway by acting on the Bcl-2 apoptosis pathway. |
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