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目的研究槲皮素(quercetin,Que)在人乳腺癌细胞系(MCF-7)死亡中的作用,初步探讨自噬和凋亡机制在MCF-7死亡中的相关性。方法采用噻唑蓝(MTT)法测定Que对MCF-7细胞的抑制作用;AO/EB染色、Hochest33258染色、单丹磺酰尸胺(MDC)、免疫荧光法观察给药后自噬和凋亡的发生;MTT法结合乳酸脱氢酶(LDH)漏出率和流式细胞仪研究自噬与凋亡机制在MCF-7死亡中的关系。结果Que对MCF-7生长有显著抑制作用,呈明显的时间、剂量依赖性;Que可诱导MCF-7细胞发生自噬和凋亡;在Que给药前用3-甲基腺嘌呤(3-MA)阻断自噬或用氯喹碱化溶酶体,Que对MCF-7的细胞毒性作用增强;碱化溶酶体后,Que可使MCF-7细胞的凋亡峰提前;溶酶体组织蛋白酶抑制剂E64d能降低Que对MCF-7细胞的生长抑制。结论Que能明显抑制MCF-7细胞的生长,并诱导其发生自噬和凋亡,自噬在早期起保护作用,另一方面溶酶体组织蛋白酶可能参与了Que诱导的MCF-7细胞死亡;溶酶体可能是MCF-7细胞发生自噬和凋亡的枢纽。
Objective To investigate the role of quercetin (Que) in the death of human breast cancer cell line (MCF-7) and to investigate the relationship between autophagy and apoptosis in MCF-7 cell death. Methods The inhibitory effect of Que on MCF-7 cells was determined by MTT assay. The apoptosis of MCF-7 cells was detected by AO / EB staining, Hochest33258 staining, MDS and immunofluorescence staining MTT assay was used to study the relationship between autophagy and apoptosis mechanism in MCF-7 death by combining the leakage rate of lactate dehydrogenase (LDH) and flow cytometry. Results Que could significantly inhibit the growth of MCF-7 cells in a time-and dose-dependent manner. Que could induce autophagy and apoptosis in MCF-7 cells. Que treated with 3-methyladenine (3- MA), cytotoxicity of Que to MCF-7 was enhanced by blocking autophagy or lysing the lysosomes with chloroquine. After alkalizing lysosomes, Que could induce apoptosis peak of MCF-7 cells in advance; lysosomes The protease inhibitor E64d reduced the inhibitory effect of Que on the growth of MCF-7 cells. Conclusions Que can significantly inhibit the growth of MCF-7 cells and induce autophagy and apoptosis. Autophagy plays a protective role in the early stage. On the other hand, lysosomal cathepsins may be involved in Que-induced MCF-7 cell death. Lysosomes may be the hub of autophagy and apoptosis in MCF-7 cells.