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[咪达唑仑对套细胞淋巴瘤JeKo-1细胞系的作用及其相关机制]

[Effect of midazolam on mantle cell lymphoma JeKo-1 cell line and its relevant mechanisms].

作者信息

Hong Jin-Quan, Wu Shan-Hu, Chen Zhi-Yuan, Zhuang Wei-Huang, Gao Hong-Zhi

机构信息

Department of Anesthesia, The Second Affiliated Hospital of Fujian Medical University, Quanzhou 362000, Fujian Province, China.

Department of Hematology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou 362000, Fujian Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Dec;21(6):1460-3. doi: 10.7534/j.issn.1009-2137.2013.06.017.

Abstract

This study was aimed to explore the effect of midazolam on mantle cell lymphoma cell line JeKo-1 and the relevant mechanisms. Effects of midazolam on the proliferation and apoptosis of JeKo-1 cells were observed by CCK8 assay and flow cytometry, respectively. Effect of midazolam on the expression of BCL-2, cytochrome C (Cyto-C), pro-caspase-9, pro-caspase-8 and pro-caspase-3 protein were detected by Western blot. The results showed that midazolam could inhibit the growth of JeKo-1 cells significantly and the concentration of 50% growth inhibition (IC50) at 48 hours was approximately 40 µmol/L. After treatment with 20, 40, 80 µmol/L midazolam for 48 hours, a dose-dependent apoptosis of JeKo-1 cells could be observed. Meanwhile, a dose-dependent reduction of BCL-2, pro-caspase-9 and pro-caspase-3 protein expression and increase of Cyto-C protein expression in JeKo-1 cells were found, but the expression of pro-caspase-8 protein did not change. It is concluded that midazolam possibly initiates the mitochondrial pathway, not the death receptor pathway, by reducing the expression of BCL-2, leading in turn to the releasing of Cyto-C in mitochondria, then activating caspase-9 and caspase-3 protein, triggers the caspase cascade, and induces the apoptosis of JeKo-1 cells ultimately.

摘要

本研究旨在探讨咪达唑仑对套细胞淋巴瘤细胞系JeKo-1的作用及其相关机制。分别采用CCK8法和流式细胞术观察咪达唑仑对JeKo-1细胞增殖和凋亡的影响。采用蛋白质免疫印迹法检测咪达唑仑对BCL-2、细胞色素C(Cyto-C)、前半胱天冬酶-9、前半胱天冬酶-8和前半胱天冬酶-3蛋白表达的影响。结果显示,咪达唑仑可显著抑制JeKo-1细胞的生长,48小时的50%生长抑制浓度(IC50)约为40µmol/L。用20、40、80µmol/L咪达唑仑处理48小时后,可观察到JeKo-1细胞呈剂量依赖性凋亡。同时,发现JeKo-1细胞中BCL-2、前半胱天冬酶-9和前半胱天冬酶-3蛋白表达呈剂量依赖性降低,Cyto-C蛋白表达增加,但前半胱天冬酶-8蛋白表达未发生变化。结论是,咪达唑仑可能通过降低BCL-2的表达启动线粒体途径,而非死亡受体途径,进而导致线粒体中Cyto-C的释放,然后激活半胱天冬酶-9和半胱天冬酶-3蛋白,触发半胱天冬酶级联反应,最终诱导JeKo-1细胞凋亡。

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