吴茱萸碱对肺癌荷瘤小鼠生长及HIF-1α/VEGF信号通路的影响

Evodiamine Inhibits the Growth of Lung Tumors in Mice via HIF-1α/VEGF Signaling Pathway

目的:探讨吴茱萸碱对Lewis肺癌荷瘤小鼠肿瘤生长及缺氧诱导因子-1α/血管内皮生长因子(HIF-1α/VEGF)信号通路的影响。方法:通过皮下注射Lewis肺癌细胞建立荷瘤小鼠模型,建模后分为模型对照组、阳性对照组(顺铂干预)及吴茱萸碱低、高剂量组,共干预14 d。通过检测各组小鼠造模给药期间肿瘤大小,计算抑瘤率,评价吴茱萸碱对肺癌荷瘤小鼠肿瘤生长的影响。此外,运用定量PCR(qPCR)法检测吴茱萸碱干预14 d后各组小鼠肿瘤组织中HIF-1α及VEGF mRNA表达的影响,免疫组织化学染色评估肿瘤组织中HIF-1α、VEGF及血小板-内皮细胞黏附分子(CD31)表达水平,计算肿瘤组织中微血管密度(MVD)。结果:吴茱萸碱对肺癌荷瘤小鼠肿瘤生长具有显著的抑制作用,阳性对照组及吴茱萸碱低、高剂量抑瘤率分别为53.13%、20.49%、38.54%;吴茱萸碱各给药剂量组可不同程度下调Lewis肺癌荷瘤小鼠肿瘤组织HIF-1α及VEGF基因及蛋白表达(P<0.01),减少肿瘤组织中MVD(P<0.01)。结论:吴茱萸碱可抑制肺癌荷瘤小鼠肿瘤生长,其作用机制可能是通过减少肿瘤组织中HIF-1α及VEGF表达,抑制肿瘤中血管生成。

Objective:To investigate the modulatory effects of evodiamine on hypoxia-inducible factor-1 alpha(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway in Lewis tumor-bearing mouse model.Methods:The tumor-bearing mouse model was established by subcutaneous injection of Lewis lung cancer cells.Then,the mice were randomized into model,positive control(cisplatin),low-dose evodiamine,and high-dose evodiamine groups and treated with corresponding agents for 14 days.The tumor size in each group was measured to calculate the tumor inhibition rate,which reflected the inhibitory effect of evodiamine on tumor growth.Furthermore,qPCR was employed to examine the mRNA levels of HIF-1αand VEGF in the tumor tissue,and immunohistochemical staining to determine the protein levels of HIF-1α,VEGF,and platelet/endothelial cell adhesion molecule-1(CD31)in the tumor tissue.The microvessel density(MVD)in the tumor tissue was calculated.Results:Evodiamine significantly decreased the tumor size in Lewis tumor-bearing mouse model.The tumor inhibitions rate in the positive control,low-dose evodiamine,and high-dose evodiamine groups were 53.13%,20.49%,and 38.54%,respectively.Evodiamine down-regulated the mRNA and protein levels of HIF-1αand VEGF and decreased the MVD in the tumor tissue(P<0.01).Conclusion:Evodiamine could inhibit angiogenesis by down-regulating the expression of HIF-1αand VEGF to hinder the growth of lung tumor in tumor-bearing mice.