bFGF抗体对人结直肠癌相关内皮细胞增殖、迁移、血管新生的影响和机制研究

Effect and mechanism of bFGF antibody on proliferation,migration and angiogenesis of human colorectal cancer associated endothelial cells

目的探究碱性成纤维细胞生长因子(bFGF)抗体对人结直肠癌相关内皮细胞增殖、迁移、血管新生及转录激活因子3(STAT3)通路的影响。方法以结直肠癌LOVO细胞培养上清诱导人脐静脉内皮(HUVECs)细胞形成的结直肠癌相关内皮细胞为研究对象。采用CCK-8法和成管实验测定bFGF抗体对bFGF刺激内皮细胞增殖、迁移、成管功能的影响。采用Western blotting法测定STAT3通路蛋白表达。结果与抗体组比较,不同浓度联合组的内皮细胞增殖抑制率均明显升高(t分别=13.65、25.95、55.07、32.43,P均<0.05);与对照组比较,不同浓度抗体组内皮细胞迁移距离明显缩短(t分别=3.69、7.19、7.29、14.34,P均<0.05),且各组抑制率升高和迁移距离缩短均呈剂量依赖性(t分别=19.57、18.00、5.59;3.51、3.46、4.77,P均<0.05)。与对照组比较,抗体组、雷抗组、联合组内皮细胞管腔形成数目均明显减少(t分别=2.58、13.85、17.16,P均<0.05),雷抗组和联合组数量明显少于抗体组,联合组明显少于雷抗组(t分别=3.14、6.15、6.67,P均<0.05)。与bFGF组比较,不同浓度抗体组细胞中pSTAT3蛋白相对表达量均明显下降(t分别=2.21、6.14、10.60、14.16,P均<0.05),且下降随着抗体浓度升高而降低(t分别=3.59、3.41、6.23,P均<0.05)。结论bFGF抗体对bFGF刺激的细胞增殖、迁移及血管新生等功效具有明显抑制作用,抑制作用存在剂量效应,且可能与抑制STAT3磷酸化有关。

Objective To investigate the effects of basic fibroblast growth factor(bFGF)antibody on the proliferation,migration,angiogenesis and transcriptional activator 3(STAT3)pathway of human colorectal cancer(CRC)related endothelial cells.Methods The formation of human umbilical vein endothelial(HUVECs)cells induced by LOVO cell culture of colorectal cancer was studied.The effects of bFGF antibody on proliferation,migration and tube formation of endothelial cells stimulated by bFGF were determined by CCK-8 method,and tube formation test.The expression of STAT3 pathway protein was determined by Western blotting.Results Compared with antibody group,the proliferation inhibition rate of endothelial cells in combined group with different concentration of bFGF antibody was significantly increased(t=13.65,25.95,55.07,32.43,P<0.05).Compared with the control group,the migration distance of endothelial cells in different antibody concentration groups was significantly shortened(t=3.69,7.19,7.29,14.34,P<0.05),and the increase of inhibition rate and the reduction of migration distance in each group were dose-dependent(t=19.57,18.00,5.59,3.51,3.46,4.77,P<0.05).Compared with the control group,the number of endothelial cell lumen formation in antibody group,thunder reactant group and combined group was significantly reduced(t=2.58,13.85,17.16,P<0.05),the number of thunder reactant group and combined group was significantly less than that in antibody group,and the number of combined group was significantly less than that in combined group(t=3.14,6.15,6.67,P<0.05).Compared with bFGF group,the expression level of pSTAT3 in all antibody groups was significantly decreased(t=2.21,6.14,10.60,14.16,P<0.05),and the decrease was significant with the increase of antibody concentration(t=3.59,3.41,6.23,P<0.05).Conlusion bFGF antibody can significantly inhibit cell proliferation,migration,and angiogenesis stimulated by bFGF.The inhibition has a dose-effect and may be related to the inhibition of STAT3 phosphorylation.