氧化槐果碱抑制Nrf2/HO-1/NQO1通路诱导乳腺癌细胞凋亡

doi:10.3870/j.issn.1672-8009.2025.06.006

摘要/Abstract

摘要： 目的探究氧化槐果碱通过调节核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase-1,HO-1)/醌氧化还原酶1[NAD(P)H:quinone oxidoreductase 1,NQO1]通路对乳腺癌(breast cancer,BC)细胞增殖、凋亡及氧化应激的影响。方法使用MDA-MB-231和MCF-7细胞系。通过CCK-8检测细胞活性;定量实时聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)和蛋白质印迹检测增殖(Survivin,Ki67,P21)、凋亡(cleaved caspase-3/9,Bax/Bcl-2)及Nrf2/HO-1/NQO1通路相关分子表达;流式细胞术分析凋亡及线粒体膜电位;酶联免疫吸附检测(ELISA)测定氧化应激指标超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)。在MDA-MB-231细胞中过表达Nrf2验证通路作用。建立裸鼠移植瘤模型评估肿瘤生长、Ki67表达、caspase-3阳性率、凋亡率(TUNEL)及Nrf2/HO-1/NQO1通路蛋白质表达。结果氧化槐果碱剂量依赖性抑制BC细胞活性,诱导凋亡,降低线粒体膜电位。其显著抑制增殖相关分子(Survivin,Ki67)表达,促进凋亡相关分子(P21,cleaved caspase-3/9,Bax/Bcl-2比值)表达;同时抑制Nrf2/HO-1/NQO1通路活化,加剧氧化应激(降低SOD、GSH,升高MDA)。过表达Nrf2可部分逆转氧化槐果碱的上述效应。动物实验证实氧化槐果碱抑制肿瘤生长、降低增殖(Ki67)、促进凋亡(caspase-3,TUNEL)并抑制体内Nrf2通路活化。结论氧化槐果碱通过抑制Nrf2/HO-1/NQO1信号通路,有效抑制乳腺癌细胞增殖、诱导凋亡并增强氧化应激。

关键词: 氧化槐果碱, 乳腺癌, 凋亡, 氧化应激, Nrf2/HO-1/NQO1信号通路

Abstract: Objective To investigate the effect of oxysophocarpine on breast cancer(BC)cell proliferation,apoptosis,and oxidative stress by modulating the nuclear factor erythroid 2-related factor 2,(Nrf2)/ heme oxygenase-1(HO-1)/ NAD(P)H:quinone oxidoreductase 1(NQO1)pathway. Methods MDA-MB-231 and MCF-7 cell lines were used.Cell viability was detected by CCK-8 assay.Quantitative real-time polymerase chain reaction(qRT-PCR)or Western blotting were used to measure the expression level of proliferation-related(Survivin,Ki67,P21),apoptosis-related(cleaved caspase-3/9,Bax/Bcl-2 ratio),and Nrf2/HO-1/NQO1 pathway-related moleculars.Apoptosis and mitochondrial membrane potential were analyzed by flow cytometry.Oxidative stress markers superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA))were assessed via enzyme-linked immunosorbent assay(ELISA).Nrf2 was overexpressed in MDA-MB-231 cells to validate the Nrf2/HO-1/NQO1 pathway’s effect.A nude mouse xenograft model was established to evaluate tumor growth,Ki67 expression,caspase-3 positivity,apoptosis rate(TUNEL),and Nrf2/HO-1/NQO1 pathway protein expression levels. Results Oxysophocarpine inhibited BC cell viability,induced apoptosis,and reduced mitochondrial membrane potential in a dose-dependent manner.It significantly suppressed the expression of proliferation-related molecules(Survivin,Ki67),and promoted the expression of apoptosis-related molecules(P21,cleaved caspase-3/9,Bax/Bcl-2 ratio).Additionally,it inhibited Nrf2/HO-1/NQO1 pathway activation and exacerbated oxidative stress(reduced SOD,GSH;increased MDA).Nrf2 overexpression partially reversed these effects of oxysophocarpine.Animal experiments confirmed that oxysophocarpine inhibited tumor growth,reduced proliferation(Ki67),promoted apoptosis(caspase-3,TUNEL),and suppressed Nrf2 pathway activation in vivo. Conclusion Oxysophocarpine effectively inhibits breast cancer cell proliferation,induces apoptosis,and enhances oxidative stress by suppressing the Nrf2/HO-1/NQO1 signaling pathway.

Key words: oxysophocarpine, breast cancer, apoptosis, oxidative stress, Nrf2/HO-1/NQO1 signaling pathway

中图分类号:

R965

崔敏, 张彤, 司梦圆, 林莹, 谢文杰, 孙吉瑞. 氧化槐果碱抑制Nrf2/HO-1/NQO1通路诱导乳腺癌细胞凋亡[J]. 医学分子生物学杂志, 2025, 22(6): 571-578.

CUI Min, ZHANG Tong, SI Mengyuan, LIN Ying, XIE Wenjie, SUN Jirui. Oxysophocarpine Induces Apoptosis of Breast Cancer Cells by Inhibition of Nrf2/HO-1 Signaling Pathway[J]. Journal of Medical Molecular Biology, 2025, 22(6): 571-578.

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