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长叶卡林A诱导多发性骨髓瘤H929细胞凋亡

[Longikaurin A induces apoptosis of multiple myeloma H929 cells].

作者信息

Zhao Shan, Pu Jian-Xin, Sun Han-Dong, Wu Ying-Li

机构信息

Department of Pathophysiology, Shanghai Jiaotong University School of Medicine, Shanghai, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2012 Jun;20(3):611-5.

Abstract

The aim of this study was to investigate the biological effect of longikaurin A on multiple myeloma H929 cells. Effects of oridonin and longikaurin A on proliferation of H929 cells were evaluated by CCK-8 assay. Cell morphological features of H929 cells were examined under inverted phase contrast microscope. Apoptosis was determined by Annexin V/PI staining. Expression of caspase-3, caspase-9, and PARP were detected by Western blot. Reactive oxygen species (ROS) level was determined by DCFDA assay. The results showed that longikaurin A (IC(50) 0.85 µmol/L) was more effective than oridonin ((IC(50) 10.66 µmol/L) to inhibit the proliferation of H929 cells. Treatment with longikaurin 2 µmol/L significantly increased the percentage of Annexin V positive cells. Caspase-3 and caspase-9 were activated and PARP, one substrate of caspase-3, was cleaved into 85 kDa fragments. The ROS scavenger N-acetyl-cysteine significantly blocked apoptosis induced by longikaurin A. However, longikaurin A did not increase the ROS level in H929 cells. It is concluded that longikaurin A is more effective than oridonin to induce apoptosis in multiple myeloma H929 cells without increasing the ROS level.

摘要

本研究旨在探讨长叶九里香素A对多发性骨髓瘤H929细胞的生物学效应。采用CCK-8法评估冬凌草甲素和长叶九里香素A对H929细胞增殖的影响。在倒置相差显微镜下观察H929细胞的形态特征。通过Annexin V/PI染色测定细胞凋亡情况。采用蛋白质免疫印迹法检测caspase-3、caspase-9和PARP的表达。通过DCFDA法测定活性氧(ROS)水平。结果显示,长叶九里香素A(IC50为0.85 μmol/L)比冬凌草甲素(IC50为10.66 μmol/L)更有效地抑制H929细胞的增殖。用2 μmol/L长叶九里香素处理可显著增加Annexin V阳性细胞的百分比。caspase-3和caspase-9被激活,caspase-3的一个底物PARP被切割成85 kDa的片段。ROS清除剂N-乙酰半胱氨酸显著阻断了长叶九里香素A诱导的细胞凋亡。然而,长叶九里香素A并未增加H929细胞中的ROS水平。结论是,长叶九里香素A比冬凌草甲素更有效地诱导多发性骨髓瘤H929细胞凋亡,且不增加ROS水平。

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