曲古菌素A对多发性骨髓瘤细胞增殖影响的相关研究

The Effets of TSA in Rgulating Cell Proliferation and Apoptosis of Multiple Myeloma KM3 Cells

目的:研究VEGF受体(Vascular endothelial growth factor receptor,VEGFR)在多发性骨髓瘤KM3细胞株中的表达及曲古菌素A(Trichostatin A,TSA)对KM3细胞增殖的影响,并进一步探讨其抗肿瘤机制。方法:体外培养KM3细胞株,MTT检测细胞增殖,流式细胞术检测细胞凋亡,RT-PCR检测VEGFR m RNA表达。结果:TSA能明显抑制KM3细胞的增殖,且具有时间剂量依赖性(P<0.05)的特点;TSA处理KM3细胞48h可诱导KM3细胞凋亡,凋亡率最高达42.72%±4.61%,呈剂量依赖性(P<0.05)特点;RT-PCR结果显示,KM3细胞仅表达VEGFR-1(flt-1),TSA能在m RNA水平抑制VEGFR-1的表达。结论:TSA可通过下调骨髓瘤细胞株KM3细胞表面VEGFR的表达,从而诱导KM3细胞的凋亡,而达到抑制其增殖的目的。

Objective:Despite of the recent advances in the treatment of multiple myeloma(MM),it is still an incurable disease in the majority of patients.In the bone marrow microenvironment of MM,VEGF is essential for tumor growth and survival,and myeloma cells are known to produce VEGF and express VEGF receptors.To investigate the effects of TSA(trichostatin A,TSA)in regulating VEGFR(vascular endothelial growth factor receptor)expression in multiple myeloma KM3cells,and study the antiproliferation mechanism of TSA on KM3cells.Methods:The cell proliferation was measured by MTT assay.The apoptosis rates were determined by flow cytometry.The mRNA level of VEGFR was determined by RT-PCR;immunocytochemistry was used to detect the protein level of VEGFR.Results:The proliferation of KM3cells decreased in TSA-treated groups in a time-dose dependent manner(P<0.05).Treatment with TSA(400、200、100and50ng·mL-1)for48h,the apoptosis rates of KM3cells were(11.77±4.64)%、(22.13±1.20)%、(23.95±.2.57)%and(42.72±4.61)%respectively.Only VEGFR-1(flt-1)was detected in KM3cells by RT-PCR and it was decreased in TSA-treated groups in a dose-dependent manner(P<0.05).Conclusion:TSA plays an important role in regulating proliferation and apoptosis of multiple myeloma cell line KM3cells,and this effect may be partly caused by decreasing the expression of VEGFR in KM3cells.