摘要
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 study the effects of qereetin（Que） -induced death of human breast cancer cell line MCF-7, and the relations between the mechanism of autophagy and apoptosis on death of MCF-7 cells. METHODS After treatment with different con- centration of Que, the growth inhibition of the MCF-7 cells was assessed by MTT colorimetric assay. The AO/EB, Hochest33258, MDC and fluorescent staining were applied to identify the autophagy and the apoptosis after Que treatment. The MTT assay, LDH leakage and the flow cytomctry were performed to analyze the relations between autophagy and apoptasis on death of MCF-7 cells. RESULTS The proliferation af MCF-7 ceils was significantly inhibited on a dose and time-dependent manner after Que treatment. Apoptosis and autophagy were induced in MCF-7 cells and detected by AO/EB, Hochest 33258, MDC and fluorescent staining in the late phase of Que treatment. With 3-MA to inhibit autophagy and Chloroquine to alkalify lysosome , the cytotoxicity of Que-induced MCF- 7 cells were enhanced. After lysosome was alkalinizied, the apoptosis peak of MCF-7 cells was advanced. E64d （ inhibitor of cathepsins） reduced the growth inhibition of Que on MCF-7 cells. CONCLUSION Que can significantly inhibit the growth of the MCF-7 ceils, and induce the autophagy and apoptosis. In the process of Que-induced death of MCF-7 cells, autophagic mechanism may play a protective role at the initiation phase. On the other hand, the lysosomal protease might take part in the death of MCF-7 cells. Therefore, in MCF-7 cell, the lysosome might be a hinge of the autophagy and the apoptosis.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2010年第6期434-439,共6页
Chinese Pharmaceutical Journal
基金
国家自然基金资助项目(30873052)
苏州大学医学发展基金(EE134703)
关键词
槲皮素
自噬
凋亡
溶酶体
quercetin
autophagy
apoptosis
lysosome
