沙利度胺对AnnexinⅡ基因的调节作用  

Effects of thalidomide on Annexin Ⅱ gene regulation

作　　者：包红雨[1] 江淼[1] 沈飞[1] 朱明清[1] 阮长耿[1] 

机构地区：[1]苏州大学附属第一医院、江苏省血液研究所,卫生部血栓与止血重点实验室,215006

出　　处：《中华血液学杂志》2009年第7期464-467,共4页Chinese Journal of Hematology

基　　金：基金项目：江苏省“135”重点学科基金

摘　　要：目的了解沙利度胺存体外对人多发性骨髓瘤（MM）细胞系RPMI8226、人微血管内皮细胞系HMEC-1中AnnexinⅡ（AnxA2）基因表达的调节作用，探讨沙利度胺诱发血栓的可能机制。方法实时荧光定量PCR检测在不同浓度下沙利度胺对两种细胞系AnxA2 mRNA表达的影响，采用流式细胞术、激光共聚焦仪检测不同浓度下沙利度胺对细胞膜表面AnxA2蛋白表达的影响。结果当沙利度胺浓度为12．5、25．0、50．0μg／ml时，RPMI8226细胞AnxA2 mRNA表达水平为0．60±0．15、0．33±0．14、0．42±0．16，较对照组（1．07±0．16）降低（P〈0．05），HMEC-1细胞AnxA2 mRNA表达水平为0．21±0．20、0．08±0．08、0．17±0．16，较对照组（1．16±0．24）降低（P〈0．05）。沙利度胺浓度为12．5、25．0、50．0μg／ml时，RPMI8226细胞膜表面AnxA2蛋白表达水平为3．39±0．32、2．82±0．28、3．21±0．23，较对照组（5．53±0．32）降低（P〈0．05），HMEC-1细胞膜表面AnxA2蛋白表达水平为0．72±0．11、0．64±0．08、0．67±0．08，较对照组（1．40±0．15）降低（P〈0．05）。结论沙利度胺能抑制RPMI8226和HMEC-1细胞AnxA2 mRNA和蛋白表达，可能为其诱发骨髓瘤血栓形成的原因之一。Objective To investigate the effect of thalidomide on Annexin Ⅱ（ AnxA2 ） gene regulation in multiple myeloma cell line RPMI8226 and human microvascular endothelial cell line HMEC-1 cells in vitro, and explore the potential mechanism of thrombosis induced by thalidomide. Methods RPMI8226 and HMEC-1 cells were cultivated in vitro. Real time quantitative PCR （RQ-PCR） was used to detect the influence of thalidomide at different concentration on the expression of AnxA2 mRNA, flow cytometry（FCM） and confocal microscopy were used to detect the cell surface protein level after the samples were stimulated with different concentrations of thalidomide. Results AnnA2 mRNA level in RPMI8226 cells treated with thalidomide at 12.5 μg/ml, 25.0 μg/ml and 50.0 μg/ml was decreased compared with the control group （0.60 ± 0.15, 0.33 ± 0. 14, 0.42 ±0. 16, vs 1.07±0. 16, respectively, P 〈0.05） and did so in HMEC-1 cells （0.21 ± 0.20, 0. 08 ± 0.08, 0.17 ± 0.16 vs 1.16 ±0.24, respectively, P 〈 0.05 ）. The AnxA2 protein level in RPMI8226 cells treated with above mentioned concentrations of thalidomide was also decreased compared with the control （3.39± 0.32, 2.82 ±0.28, 3.21±0.23 vs 5.53 ±0.32, repectively, P 〈 0.05 ） and that did so in HMEC-1 cells （0.72±0.11,0.64 ±0.08, 0.67±0.08 vs 1.40 ±0.15, respectively, P 〈0.05）. Conclusions Thalidomide can inhibit the expression of AnxA2 mRNA and protein in RPMI8226 and HMEC-1 cells, which may be one of the mechanisms for the development of thrombosis induced by thalidomide in multiple myeloma patients.

关 键 词：沙利度胺 AnnexinⅡ RPMI8226细胞 HMEC-1细胞 血栓形成 

分 类 号：R686[医药卫生—骨科学]