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目的:探讨可溶性Tie2(soluble Tie 2,sTie2)对结肠癌HCT116细胞血管生成拟态(vascular mimicry,VM)形成、增殖、迁移及侵袭能力的影响。方法:将重组质粒pBLAST49-hsTie2及对照质粒pBLAST49通过脂质体转染至HCT116细胞,分别形成hsTie2-HCT116细胞和Ctrl-HCT116细胞。通过3D模型培养、SRB法、细胞划痕实验及Transwell法分别检测HCT116细胞的VM形成、增殖、迁移及侵袭能力,采用Western blotting法检测HCT116细胞中VE-cadherin蛋白的表达。结果:pBLAST49-hsTie2重组质粒成功转染至结肠癌HCT116细胞。与Ctrl-HCT116细胞相比,hsTie2-HCT116细胞中VM的形成[(0.75±0.45)vs(7.50±0.52)个/视野,P<0.01]及VE-cadherin蛋白的表达[(1.23±0.08)vs(1.73±0.02),P<0.05]显著降低;细胞增殖率也显著降低[(32.57±4.57)%vs(88.24±21.94)%,P<0.01];细胞迁移能力[(0.37±0.07)vs(0.80±0.03)mm,P<0.01]及侵袭能力[(57.25±3.17)vs(127.25±6.25)个/视野,P<0.01]均显著减弱。结论:sTie2通过阻抑VM形成抑制结肠癌细胞的增殖、迁移和侵袭,有望成为既抗血管生成又抗VM形成的双靶向治疗结肠癌的药物。
Objective: To investigate the effect of soluble Tie2 (sTie2) on the formation, proliferation, migration and invasion of human colon cancer HCT116 cells. Methods: The recombinant plasmid pBLAST49-hsTie2 and control plasmid pBLAST49 were transfected into HCT116 cells by liposome to form hsTie2-HCT116 cells and Ctrl-HCT116 cells, respectively. The formation, proliferation, migration and invasion of HCT116 cells were detected by SRB method, cell scratch assay and Transwell method respectively. The expression of VE-cadherin protein in HCT116 cells was detected by Western blotting. Results: pBLAST49-hsTie2 recombinant plasmid was successfully transfected into colon cancer HCT116 cells. The formation of VM in hsTie2-HCT116 cells was significantly higher than that of Ctrl-HCT116 cells [(0.75 ± 0.45 vs vs (7.50 ± 0.52) / visual field, P <0.01] and the expression of VE-cadherin protein [(1.23 ± 0.08) vs (1.73 ± 0.02), P <0.05], and the cell proliferation rate was also significantly lower (32.57 ± 4.57% vs 88.24 ± 21.94%, P <0.01) 0.80 ± 0.03) mm, P <0.01] and invasive ability [(57.25 ± 3.17) vs (127.25 ± 6.25) / visual field, P <0.01]. Conclusion: sTie2 can inhibit the proliferation, migration and invasion of colon cancer cells by inhibiting the formation of VM, which is expected to become a dual-targeted therapy for colon cancer that is both anti-angiogenic and anti-VM.