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姜黄素联合硼替佐米对人多发性骨髓瘤细胞系H929增殖及凋亡的影响及其机制

[Effect of curcumin in combination with bortezomib on proliferation and apoptosis of human multiple myeloma cell line H929 and its mechanism].

作者信息

Zhang Xiao-Yan, Bai Qing-Xian, Huang Gao-Sheng, Zhao Hui, Chen Juan-Juan, Yang Li-Jie

机构信息

Department of Hematology, Xijing Hospital, Xi'an 710032, Shaanxi Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Jun;19(3):684-8.

Abstract

This study was aimed to investigate the effect of curcumin in combination with bortezomib on the proliferation and apoptosis of human MM cell line H929 in vitro, and to explore its mechanisms. MTT assay was applied to detect the inhibitory effects of curcumin and bortezomib either alone or combined at different concentrations on H929 cells, and flow cytometry was employed to assay the apoptosis rate. In addition, RT-PCR was used to analyze the mRNA expression of gene BCL-2, BAX, cyclin D1. Immunofluorescence technique was performed to study the location changes of NF-κB P65 in different groups. The results showed that both curcumin and bortezomib inhibited the proliferation of MM cell line H929 in dose-dependent manner, and combination of these two drugs displayed synergistical effect. A much higher apoptosis rate was determined by flow cytometry in combinative groups than that in single or control group. And RT-PCR showed, as compared with curcumin or bortezomib group, there was mRNA expression decrease of BCL-2, cyclin D1 but increase of BAX in combined group. The expression of NF-κB P65 in nucleus was downregulated in either the curcumin or bortezomib group, however, distribution of NF-κB P65 in cytoplasm was observed in combined group. It is concluded that the combination of curcumin and bortezomib is much more effective for the inhibiting proliferation and promoting apoptosis of H929 cell line, which may function by inhibiting the transcription of NF-κB and apoptosis-related genes.

摘要

本研究旨在探讨姜黄素联合硼替佐米对人骨髓瘤细胞系H929体外增殖及凋亡的影响,并探究其作用机制。采用MTT法检测不同浓度的姜黄素、硼替佐米单独及联合应用时对H929细胞的抑制作用,运用流式细胞术检测细胞凋亡率。此外,采用RT-PCR分析基因BCL-2、BAX、细胞周期蛋白D1的mRNA表达。运用免疫荧光技术研究不同组中NF-κB P65的定位变化。结果显示,姜黄素和硼替佐米均以剂量依赖方式抑制骨髓瘤细胞系H929的增殖,二者联合应用表现出协同效应。流式细胞术检测结果表明,联合用药组的凋亡率显著高于单药组和对照组。RT-PCR结果显示,与姜黄素组或硼替佐米组相比,联合用药组中BCL-2、细胞周期蛋白D1的mRNA表达降低,而BAX的表达升高。姜黄素组和硼替佐米组细胞核中NF-κB P65的表达均下调,而联合用药组中观察到NF-κB P65在细胞质中的分布。结论:姜黄素联合硼替佐米对抑制H929细胞系的增殖和促进其凋亡更有效,其作用机制可能是通过抑制NF-κB及凋亡相关基因的转录实现的。

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