摘要
目的 研究自噬在槲皮素（quercetin，Que）诱导的人肝癌细胞系SMMC-7721死亡中的作用。方法 采用四甲基偶氮唑蓝法测定槲皮素对SMMC-7721细胞的抑制作用；单丹磺酰戊二胺、免疫荧光法观察给药后自噬的发生； 乳酸脱氢酶 (LDH)漏出率、蛋白印迹法观察槲皮素诱导的SMMC-7721细胞死亡的分子机制。结果 槲皮素对SMMC-7721生长有显著抑制作用，呈明显的时间、剂量依赖性； 槲皮素可激活SMMC-7721 细胞的LC3的表达、诱导自噬的发生；乳酸脱氢酶漏出率实验结果表明，在槲皮素给药前用3-甲基腺嘌呤（3-MA）阻断自噬或者氯喹碱化溶酶体，可增强槲皮素对SMMC-7721细胞的细胞毒作用； 蛋白印迹结果显示，槲皮素可使SMMC-7721细胞溶酶体组织蛋白酶B的表达明显增强。结论 槲皮素能明显抑制SMMC-7721 细胞的生长，并诱导其发生自噬；自噬激活能降低槲皮素对SMMC-7721细胞的毒性；自噬的发生与溶酶体酶组织蛋白酶B表达增强相关。
Abstract
OBJECTIVE A im To investigate the role of qercetin( Que)-induced autophagy in the death of hepatocellular carcinoma cell line SMMC-7721 cells. METHODS After treatment with different concentration of Que， the growth inhibition of the SMMC-7721 cells were assessed by MTT colorimetric assay. The fluorescent staining was applied to identify the autophagy after Que treatment. Lactate dehydrogenase (LDH) leakage and Western blot analysis were used to study the autophagic mechanisms involved in death of SMMC-7721 cells. RESULTS The proliferation of SMMC-7721 cells were significantly inhibited in a dose and time-dependent manner after Que treatment. Autophagy was also induced in MCF-7 cells as detected by MDC staining and Fluorescent staining in the early phase. The autophagy specific inhibitor 3-MA or chloroquine potentiated Que′s cytotoxicitiy in SMMC-7721 cells when administered 1h before Que； The expression of cathepsin B increased after Que treatmen. CONCLUSION Que can significantly inhibit the growth of the SMMC-7721 cells by inducing the autophagy， which is a protection mechanism that can reduce the cytotoxicity induced by Que in SMMC-7721 cells. The activation of Cathepsin B is considerable in autophagy process.
关键词
槲皮素 /
自噬 /
LC3 /
溶酶体
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Key words
quercetin /
autophagy /
LC3 /
lysosome
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朱茉莉a， b，梁金英a，李鹏a，赵繁荣a，张玉林 a，高志涛c*.
自噬对槲皮素诱导人肝癌SMMC-7721细胞死亡的影响[J]. 中国药学杂志, 2012, 47(13): 1052-1055
ZHU Mo-lia，b，LIANG Jin-yinga ， LI Penga， ZHAO Fan-ronga， ZHANG Yu-lina ， GAO Zhi-taoc*(a..
Effects of Autophagy on Quercetin-Induced Death of SMMC-7721 Cell[J]. Chinese Pharmaceutical Journal, 2012, 47(13): 1052-1055
中图分类号：
R965
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参考文献
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脚注
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基金
河南省药理学重点学科开放性课题（ZD200906）
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