MCP-1转染人脐静脉内皮细胞过表达/沉默后相关信号通路研究

The Study of Signaling Pathways in MCP-1 Over-Expression / Interference of HUVECs

目的探讨单核细胞趋化蛋白-1(MCP-1)转染人脐静脉内皮细胞过表达/沉默后相关信号通路与深静脉血栓形成的关系。方法培养人脐静脉内皮细胞行免疫荧光、免疫共沉淀检测,构建人MCP-1细胞系过表达/沉默载体,基因表达谱芯片检测人MCP-1细胞系过表达/沉默后转录谱变化并行生物信息技术分析。结果培养人脐静脉内皮细胞;构建人MCP-1过表达/干扰载体MCP-1-p CDH-GFP/MCP-1-LMP sh RNAmir1经病毒转染后感染HUVECs;基因芯片检测发现与正常细胞相比,过表达载体MCP-1-p CDH-GFP转染细胞有18条信号通路下调,7条信号通路上调;干扰载体MCP-1-LMP sh RNAmir1转染细胞有60条信号通路下调,15条信号通路上调。结论MCP-1转染人脐静脉内皮细胞后相关信号通路为深静脉血栓疾病分子层面研究提供新的思路,MCP-1可能在深静脉血栓形成发生发展过程中发挥重要作用。

Objective To investigate the association between the signaling pathways of MCP-1-pCDH-GFP-transfected cells and deep venous thrombosis （DVT）. Methods The cultured human umbilical vein endothelial cells （HUVECs） were tested by immunofluorescence and co-immunoprecipitation methods. The constructed MCP-1 over-expression/interference vector, and the change of transcription profile were detected by microarray assay and biological information technology analysis. Results MCP-1 over-expression/interference vector MCP-1-pCDH-GFP/MCP-1-LMP shRNAmir1 was con？structed and HUVECs were transfected. According to the microarray analysis we found that there were 18 down-expressed signaling pathways and 7 up-expressed signaling pathways in MCP-1-pCDH-GFP-transfected cells. There were 60 down-expressed signaling pathways and 15 up-expressed signaling pathways in the MCP-1-LMP shRNAmir1 transfected cells. Conclusion Signaling pathways of MCP-1 plays an important role in DVT formation,which may provide us a new way to study molecular mechanism of DVT.