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目的:研究缺氧对肝癌血管生成拟态(vasculogenic mimicry,VM)的影响及其可能机制.方法:建立基质胶Matrigel三维培养体系,观察高转移人肝癌细胞系MHCC97-H在常氧(常氧组)和缺氧(缺氧组)条件下形成VM的差异;应用RT-PCR和Western blot技术检测两组缺氧诱导因子-1α(HIF-1α)、基质金属蛋白酶(matrix metalloproteinase,MMP)-2和MMP-9的表达.选择151例肝癌组织标本,免疫组织化学方法检测VM是否存在及HIF-1α、MMP-2和MMP-9的表达.结果:三维培养48h,缺氧组能形成基质型VM或管形VM,而常氧组不形成基质型VM或形成较短的管形VM.缺氧组HIF-1α、MMP-2和MMP-9 mRNA和蛋白表达水平明显较常氧组高(mRNA:0.70±0.05vs0.51±0.10,P=0.03;0.90±0.09vs0.67±0.09,P=0.013;0.64±0.10vs0.42±0.07,P=0.01;蛋白:0.52±0.09vs0.41±0.09,P=0.035;0.63±0.11vs0.50±0.11,P=0.023;0.50±0.14vs0.38±0.09,P=0.008).151例肝癌组织标本中31例存在VM,存在VM的标本HIF-1α、MMP-2和MMP-9免疫组织化学染色综合得分较不存在VM的标本高(4.5±1.6vs3.4±1.7,P=0.001;4.5±1.6vs3.4±1.7,P=0.030;3.9±1.9vs2.9±1.8,P=0.007).结论:缺氧能促进肝癌VM的形成,诱导HIF-1α、MMP-2和MMP-9表达可能是其作用机制之一.
Objective: To investigate the effect of hypoxia on vasculogenic mimicry (VM) and its possible mechanism.Methods: Matrigel three-dimensional culture system was established to observe the expression of MHCC97-H in normoxia (Hypoxia group) and hypoxia group (hypoxia group). The expressions of HIF-1α, MMP-2 and MMP-9 were detected by RT-PCR and Western blot, The expression of HIF-1α, MMP-2 and MMP-9 were detected by immunohistochemical method in 151 cases of hepatocellular carcinoma.Results: In the three-dimensional culture for 48 hours, the hypoxia group could form the stroma VM or tubular VM, while the normoxic group did not form the matrix-type VM or formed a shorter tubular VM.HIF-1α, MMP-2 and MMP-9 mRNA and protein expression in hypoxia group was significantly higher than that in normoxia group mRNA: 0.70 ± 0.05 vs 0.51 ± 0.10, P = 0.03; 0.90 ± 0.09 vs. 0.67 ± 0.09, P = 0.013; 0.64 ± 0.10 vs. 0.42 ± 0.07, P = 0.01; Protein: 0.52 ± 0.09 vs. 0.41 ± 0.09, P = 0.035; 0.63 ± 0.11vs0.50 ± 0.11, P = 0.023; 0.50 ± 0.14vs0.38 ± 0.09, P = 0.008) .There were VMs in 31 cases of HCC tissues and HIF- 1α, MMP-2 and MMP-9 The scores of histological staining in the specimens were higher than those without VM (4.5 ± 1.6 vs 3.4 ± 1.7, P = 0.001; 4.5 ± 1.6 vs 3.4 ± 1.7, P = 0.030; 3.9 ± 1.9 vs 2.9 ± 1.8, P = 0.007) .Conclusion: Hypoxia can promote the formation of VM and induce the expression of HIF-1α, MMP-2 and MMP-9 in hepatocellular carcinoma.