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徐建永,许达才,洪跃辉,等.大麻受体激动剂WIN对白血病K562细胞增殖和诱导凋亡作用的研究[J].同济大学学报（医学版）,2014,35(1):15-20. [点击复制]
XU Jian-yong,XU Da-cai,HONG Yue-hui,et al.Effects of cannabinoid receptor activator WIN on proliferation and apoptosis of K562 Cells[J].同济大学学报（医学版）,2014,35(1):15-20. [点击复制]

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大麻受体激动剂WIN对白血病K562细胞增殖和诱导凋亡作用的研究
徐建永¹,许达才²,洪跃辉²,陈新美²,赵青²
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(1.广州医科大学卫生职业技术学院生物化学教研室，广东广州510925;2.广州医科大学生物化学教研室，广东广州510182)

摘要:

目的探讨大麻受体激动剂WIN-55，2d(WIN)对白血病K562细胞增殖和凋亡的作用及机制。方法将细胞分成对照组和不同剂量大麻受体激动剂WIN用药组D CCK-8测定WIN对K562细胞增殖的影响；DAPI染色观察细胞核形态变化;JC-1分析线粒体膜电位变化；分光光度法检测Caspase-3、Caspase-8、Caspase-9的治性；流式细胞仪分析细胞凋亡率变化;Western印迹法分析Bax、Bcl-2及C-myc蛋白的表达。结果与对照组相比较，5、10、20ηmol/L的WIN处理K562细胞24、48 h后，增殖抑制作用明显且具有剂量依赖性，差异有统计学意义（P< 0.05)。DAPI染色和线粒体膜电位检测结果显示，K562细胞发生凋亡。流式细胞仪检测结果显乐，细胞凋亡率升高。WIN 处理24h 后，Caspase-3、Caspase-8、Caspase-9 活性均增加（P<0.05)。Western 印迹法显示，Bax 蛋白表达增加，C-myc、Bcl-2蛋白表达下降。结论大麻受体激动剂WIN能抑制白血病K562细胞增殖，并诱导凋亡，其机制可能通过上调Bax蛋白表达，下调C-myc、Bcl-2蛋白表达，以及促使线粒体跨膜电位下降，激活Caspase-3、 Caspase-8、Caspase-9 蛋白而实现。

关键词: WIN-55，2〗2-2 K562细胞；凋亡；增殖

DOI：10.3969/j.issnl008-0392.2014.01.004

基金项目:国家自然科学基金（SIMUOl);国家自然科学基金（81372466);广州市属高校科研计划青年项目（2012C118)

Effects of cannabinoid receptor activator WIN on proliferation and apoptosis of K562 Cells

XU Jian-yong¹,XU Da-cai²,HONG Yue-hui²,CHEN Xin-mei²,ZHAO Qing²

(1. Dept. of Biochemistry, College of Health Sciences, Guangzhou Medical University, Guangzhou 510925, Guangdong Province, China;2. Dept. of Biochemistry, Guangzhou Medical University, Guangzhou 510182, Guangdong Province, China)

Abstract:

Objective To investigate the effect of cannabinoid receptor activator WIN-55,212-2 (WIN) on the proliferation and apoptosis of K562 cells, and its mechanism.Methods K562 cells were treated with 5，10 or 20ηmol/L WIN for 24 h or 48 h in vitro. Cell proliferation was measured by Cell Counting Kit-8；mitochondrial membrane potential was determined by JC-lassay kit； the apoptosis of K562 cells was examined by Cell Apoptosis DAPI Detection Kit and flow cytometry. The caspase-3 ,caspase-8, caspase-9 activity was measured by colorimetric method；the expression of C-myc’Bax and Bcl-2 was detected by Western blot assay.Results Compared to control group, the proliferation of K562 cells was inhibited by WIN treatment in a dose-dependent manner (P<0.05). DAPI staining and mitochondrial membrane potential detection showed that WIN induced apoptosis of K562 cells. Flow cytometry showed that the apoptotic rates of K562 cells increased after WIN treatment. Compared to control group, the activity of caspase-3，caspase-8 and caspase-9 was increased in WIN-treatment group (P<0.05). Western blot showed that WIN treatment resulted in down-regulation of C-myc and Bcl-2 expression and up-regulation of Bax expression. Conclusion Cannabinoid receptor activator WIN-55,212-2 can inhibit the proliferation and induce apoptosis of K562 cells, which may be associated with the activation of caspases，down-regulation of C-myc and Bcl-2 expression and up-regulation of Bax expression.

Key words: WIN-55,212-2 K562 cells apoptosis proliferation