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毛酸浆内酯诱导人乳腺癌MCF-7细胞凋亡的机制研究
余雅琴, 何昕雅, 张强, 康宁
天津中医药大学中西医结合学院,天津 301617
摘要:
[目的] 探讨毛酸浆活性成分毛酸浆内酯(PPB)诱导人乳腺癌MCF-7细胞凋亡的机制。[方法] 采用噻唑蓝法(MTT法)检测PPB对MCF-7细胞及人外周血单核细胞(hPBMC)增殖的影响;克隆形成实验考察PPB对MCF-7细胞克隆形成的影响;MTT法考察泛半胱天冬氨酸蛋白酶(Caspase)抑制剂Z-VAD-fmk对PPB诱导MCF-7细胞死亡的作用;Western Blot法考察PPB处理后凋亡相关蛋白PARP、Caspase-7/8/9、Cytochrome c、Bax、Bcl-2、Fas及FADD的表达水平。[结果] PPB时间、剂量依赖性抑制MCF-7细胞生长,且对hPBMC无明显细胞毒性;PPB呈剂量依赖性地抑制MCF-7细胞克隆形成;PPB处理后,凋亡特征性蛋白PARP、原型Caspase-7蛋白表达量显著减少,而Cleaved PARP、Cleaved Caspase-7显著增加;同时内源性凋亡途径相关蛋白Bax、Cleaved Caspase-9、Cytochrome c表达量显著上调,而Bcl-2及Caspase-9原型形式显著下调。外源性凋亡相关蛋白Fas、Cleaved Caspase-8及FADD表达量上调,而Caspase-8原型形式显著下调;加入Z-VAD-fmk可显著逆转PPB诱导的MCF-7细胞死亡。[结论] 毛酸浆活性成分PPB可以诱导MCF-7细胞发生内源性和外源性凋亡。
关键词:  乳腺癌  毛酸浆内酯  细胞凋亡
DOI:10.11656/j.issn.1673-9043.2022.04.13
分类号:R285.5
基金项目:国家自然科学基金项目(81774087)
Mechanism of inducing apoptosis in human breast cancer MCF-7 cells by Physapubesin
YU Yaqin, HE Xinya, ZHANG Qiang, KANG Ning
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Abstract:
[Objective] To investigate the anti-breast cancer effect and mechanism of Physapubesin (PPB) from Physalis pubescens L.[Methods] MTT assay was used to detect the effect of PPB on the proliferation of MCF-7 cells and the peripheral blood mononuclear cells(hPBMC). Colony formation assay was used to investigate the effect of on the colony formation of MCF-7 cells. MTT assay was used to investigate the effect of apoptosis inhibitor Z-VAD-fmk on the cell death of MCF-7 cells. The expression of apoptosis related proteins PARP, Caspase-7/8/9, cytochrome c, Bax, Bcl-2, Fas and FADD at different concentrations of PPB were detected by Western Blot.[Results] PPB inhibited the growth of MCF-7 cells in the time- and dose-dependent manners, and it has no obvious toxicity to hPBMC cells; The results of colony formation assay showed that PPB could inhibit MCF-7 cell clone formation in a dose-dependent manner; The characteristic apoptotic protein PARP, Caspase-7 was significantly reduced by PPB, while the cleaved PARP, Cleaved Caspase-7 were significantly increased. The protein of endogenous apoptotic pathway including Bax, Cleaved Caspase-9, Cytochrome c were significantly up-regulated, at the same time Bcl-2 and Caspase-9 was significantly down-regulated. In addition, the exogenous apoptotic related proteins Fas, Cleaved Caspase-8 and FADD were up-regulated by treatment with PPB, but the protein levels of Caspase-8 were down-regulated. The addition of Z-VAD-fmkcould markedly reverse the death of MCF-7 cells induced by PPB.[Conclusion] PPB can induce endogenous and exogenous apoptosis in the MCF-7 cells.
Key words:  breast cancer  Physapubesin  apoptosis
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