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去乙酰化酶抑制剂Scriptaid对骨髓瘤IM9细胞的影响及作用机制

Effects of HDAC Inhibitor Scriptaid on IM9 Cell Line and Its Mechanism
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摘要 目的:研究组蛋白去乙酰化酶抑制剂Scriptaid对骨髓瘤细胞IM9的生物学作用,并初步探讨Scriptaid诱导其凋亡的作用机制。方法:选用多发性骨髓瘤细胞IM9,应用CCK8法检测Scriptaid对IM9细胞活力的影响,流式细胞术检测Scriptaid对IM9细胞周期及凋亡的影响,应用RT-PCR技术鉴定Scriptaid诱导凋亡的相关靶基因,进一步通过报告基因检测Scriptaid对p21启动子活性的影响。结果:去乙酰化酶抑制剂Scriptaid抑制IM9细胞的活力,且呈浓度依赖性(r=0.9865)。Scriptaid能够使IM9细胞阻滞在G2/M期,伴随着药物浓度升高,G2/M期细胞比例显著增高(r=0.9845)。Scriptaid对IM9细胞凋亡有明显的促进作用,凋亡相关蛋白Caspase 9、Caspase 3以及PARP1被激活。Scriptaid处理IM9细胞后,凋亡相关因子BAD、PTEN及p21表达升高,其中p21启动子的活性被显著激活。结论:去乙酰化酶抑制剂Scriptaid可以通过转录激活p21促进IM9细胞的凋亡。 Objective: To study the effect of HDAC inhibitor Scriptaid on multiple myeloma IM9 cells and preliminarily clarify the mechanism of Scriptaid-induced cell apoptosis. Methods: The cell viability,cell cycle and cell apoptosis were measured by CCK8 assay and flow cytometry respectively,the relative target gene expression levels were detected by RT-PCR,the effect of Scriptaid on p21 promoter activity was detected by using luciferase reporter assay.Results: Scriptaid inhibited IM9 cell viability in a dose-dependent manner. Scriptaid induced IM9 cell cycle arrest at G2/M phase in a dose-dependent manner. Scriptaid triggered IM9 cell apoptosis was obviously,the mRNA levels of apoptosis-related proteins Caspase 9,Caspase 3 and PARP1 were also activated. The apoptosis-associated factors BAD,PTEN and p21 increased following treatment with different dose of Scriptaid,meanwhile,p21 promoter activity was also activated significantly. Conclusion: HDAC inhibitor Scriptaid can promote IM9 cell apoptosis by transcriptional activation of p21 promoter in concentration-dependent manner.
作者 刘琳琳 孙筱洋 谢钰 韩丹阳 姚若斯 徐开林 LIU Lin-Lin;SUN Xiao-Yang;XIE Yu;HAN Dan-Yang;YAO Ruo-Si;XU Kai-Lin(Department of Hematology,The Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,Jiangsu Province,China)
出处 《中国实验血液学杂志》 CAS CSCD 北大核心 2018年第4期1116-1121,共6页 Journal of Experimental Hematology
基金 国家自然科学基金(81600173) 江苏省自然科学基金(BK20160230)
关键词 Scriptaid 多发性骨髓瘤 IM9细胞系 细胞凋亡 P21 scripatid multiple myeloma IM9 cell line cell apoptosis p21
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