组蛋白去乙酰化酶抑制剂LBH589诱导多发性骨髓瘤细胞凋亡及其耐药机制逆转的研究

[Study on histone deacetylase inhibitor LBH589 induces apoptosis of multiple myeloma cells and its reversal of drug resistance mechanism].

作者信息

Zhang Ling, Ma Yan-ping, Jia Gu

机构信息

Department of Hematology, Second Hospital of Shanxi Medical University, Taiyuan, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2012 Nov;33(11):926-31.

PMID:23363750

Abstract

OBJECTIVE

To explore the impact of a new generation of histone deacetylase inhibitor LBH589 alone or in combination with proteasome inhibitor bortezomib on multiple myeloma (MM) cells proliferation and its mechanism.

METHODS

MM cell line U266 and dexamethasone resistant cell line MM1R cells were treated with different concentrations of LBH589 alone or in combination with bortezomib, the inhibition of cells proliferation was detected by MTT, the cell cycle and apoptosis by flow cytometry. The expression level of histone H4 acetylation and PARP, Bcl-X protein was analyzed by western blot, expression level of caspase-3, APAF-1 and TOSO gene by real-time fluorescence quantitative PCR.

RESULTS

U266 and MM1R cell proliferation were inhibited by different concentrations of LBH589 (0, 10, 20, 50 nmol/L) alone or 50 nmol/L of LBH589 in combination with bortezomib (10, 20 nmol/L) in a dose- and time-dependent manner. Inhibition effect was significantly higher in all combinative groups than in single agent groups (all P < 0.05). The percentage of G(0)/G(1) phase in MM1R cells were 36.60%, 46.50%, 51.40%, 57.10%, 75.48%, 79.73%, respectively, and the apoptosis rate were 5.27%, 31.41%, 39.78%, 44.07%, 73.60%, 83.27%, respectively. The effects appeared to occur in a dose-dependent manner, and being significantly higher in all combinative groups than in single agent groups (all P < 0.05). The expression of the caspase-3 and APAF-1 gene up-regulated gradually, while TOSO gene expression in MM1R cells down-regulated gradually in a dose- and time-dependent manner (all P < 0.05).

CONCLUSIONS

LBH589 can inhibit the growth of MM cells, block the cell cycle and induce cell apoptosis, which has anti-resistant effect on multidrug resistant cell. At the same time LBH589 in combination with bortezomib on myeloma cell has a synergistic effect, its mechanism and reversal of drug resistance mechanism involves in multiple changes in gene expression.

摘要

目的

探讨新一代组蛋白去乙酰化酶抑制剂LBH589单独或联合蛋白酶体抑制剂硼替佐米对多发性骨髓瘤（MM）细胞增殖的影响及其机制。

方法

用不同浓度的LBH589单独或联合硼替佐米处理MM细胞系U266和地塞米松耐药细胞系MM1R细胞，采用MTT法检测细胞增殖抑制率，流式细胞术检测细胞周期和凋亡情况。通过蛋白质印迹法分析组蛋白H4乙酰化及PARP、Bcl-X蛋白的表达水平，实时荧光定量PCR检测caspase-3、APAF-1和TOSO基因的表达水平。

结果

不同浓度的LBH589（0、10、20、50 nmol/L）单独或50 nmol/L的LBH589联合硼替佐米（10、20 nmol/L）均能以剂量和时间依赖性方式抑制U266和MM1R细胞的增殖。所有联合组的抑制效果均显著高于单药组（均P<0.05）。MM1R细胞G(0)/G(1)期百分比分别为36.60%、46.50%、51.40%、57.10%、75.48%、79.73%，凋亡率分别为5.27%、31.41%、39.78%、44.07%、73.60%、83.27%。这些效应呈剂量依赖性，且所有联合组均显著高于单药组（均P<0.05）。MM1R细胞中caspase-3和APAF-1基因的表达呈剂量和时间依赖性逐渐上调，而TOSO基因表达呈剂量和时间依赖性逐渐下调（均P<0.05）。