论文部分内容阅读
研究重组福安泰-03(recombinant Fuantai-03,rFAT-03)对人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)迁移和增殖的影响.聚碳酸酯膜小室趋化运动模型(transwell model)检测rFAT-03对HUVECs迁移能力的影响;MTT法检测rFAT-03对HUVECs和多种人癌细胞生长的影响;荧光显微镜观察rFAT-03作用下HUVECs的形态变化;膜联蛋白V-异硫氰酸荧光素(annexin V-fluorescein isothiocyanate,annexinV-FITC)双染检测rFAT-03对HUVECs早期凋亡的影响;流式细胞术分析rFAT-03对HUVECs周期及凋亡的影响;Western印迹检查rFAT-03对HUVECs血管内皮细胞生长因子(VEGF)、Bax和Bcl-2表达的影响.结果显示,rFAT-03明显抑制HUVECs细胞的迁移和增殖,其抑制效果与剂量和作用时间相关.0.20mg/mL恩度(endostar),0.10、0.20mg/mLrFAT-03作用HUVECs24h,对细胞迁移的抑制率分别为32.0%、32.6%、57.1%(P<0.01).0.20mg/mL恩度,0.05、0.10、0.20mg/mLrFAT-03作用HUVECs72h,其对细胞生长的抑制率分别为40.9%、63.7%、69.3%、87.0%.但rFAT-03对多种人癌细胞株的生长却无明显影响.rFAT-03处理HUVECs24h,细胞的早期凋亡率增加(P<0.05).rFAT-03阻滞HUVECs于G0/G1期,并呈现典型的凋亡峰.终浓度为0.05、0.10、0.20mg/mLrFAT-03作用于HUVECs24h,G0/G1期细胞指数分别为63.4%、67.5%和75.7%(对照组为62.1%),凋亡指数分别为4.2%、8.5%和10.3%.rFAT-03下调HUVECs的VEGF和抑调亡基因Bcl-2的表达,上调促凋亡基因Bax的表达,其效果与剂量相关.结果提示,rFAT-03明显抑制HUVECs细胞的迁移和增殖,诱导其凋亡,它的这些作用与其下调VEGF、Bcl-2表达,上调Bax表达密切相关.
To investigate the effect of recombinant Fuantai-03 (rFAT-03) on the migration and proliferation of human umbilical vein endothelial cells (HUVECs) in vitro.METHODS The transwell model The effect of rFAT-03 on the growth of HUVECs and a variety of human cancer cells was examined by MTT assay. The morphological changes of HUVECs under the action of rFAT-03 were observed by fluorescence microscopy. The effect of rFAT-03 on the early apoptosis of HUVECs was detected by double staining with annexin V-fluorescein isothiocyanate (Annexin V-FITC). The effect of rFAT-03 on the cycle and apoptosis of HUVECs was analyzed by flow cytometry. The rFAT -03 on the expression of vascular endothelial growth factor (VEGF), Bax and Bcl-2 in HUVECs.The results showed that rFAT-03 significantly inhibited the migration and proliferation of HUVECs, and its inhibitory effect was related to dose and duration of action.Of 0.20 mg / mL endostar and 0.10, 0.20mg / mL rFAT-03 for 24 h, the inhibitory rates of cell migration were 32.0%, 32.6%, 57.1% (P <0.01) , 0.20mg / mL rFAT-03 role of HUVECs 72h, its cells However, rFAT-03 had no significant effect on the growth of a variety of human cancer cell lines.After treated with rFAT-03 for 24 h, the early apoptosis rate of HUVECs increased P <0.05) .RFAT-03 blocked HUVECs in G0 / G1 phase and showed a typical apoptotic peak.The cell index of G0 / G1 phase was 0.05, 0.10, 0.20mg / mL rFAT- 63%, 67.5%, and 75.7% (62.1%) in the control group, and the apoptotic index was 4.2%, 8.5% and 10.3% respectively.rFAT-03 downregulated the expression of VEGF and down-regulated the expression of Bcl-2 in HUVECs The results showed that rFAT-03 significantly inhibited the migration and proliferation of HUVECs and induced its apoptosis, and these effects were associated with its downregulation of VEGF and Bcl-2 expression and up-regulation of Bax expression Related.