新藤黄酸干预肺腺癌血管生成的细胞学研究

摘要

目的 通过对人脐静脉内皮细胞HUVEC和人肺腺癌A549的培养,检测含新藤黄酸(GNA)条件培养基对血管内皮细胞存活率、成管和生长的影响.方法 采用甲基噻唑基四唑(MTT)法和平板克隆实验法研究GNA对HUVEC存活率和克隆形成率的影响;应用薄层胶原建立血管内皮细胞的二维培养模型,观察GN A对于血管内皮细胞成管现象的影响;采用细胞划痕愈合和小室迁移实验考察GNA对HUVEC的迁移能力影响;Westernblot检测血管内皮生长因子(VEGF)和缺氧诱导因子(HIF-1α)蛋白的表达.结果 MTT检测结果显示,HUVEC细胞存活率和克隆形成率随GNA剂量增加而降低.GNA可抑制HUVEC细胞的迁移.还可抑制HUVEC管腔样结构形成.此外,GNA可下调HUVEC中VEGF和HIF-1α蛋白的表达.结论GNA可在体外抑制血管生成,其作用机制可能与抑制肿瘤细胞分泌的HIF-1α和VEGF有关.%Objective By investigating the survival rate tubule formation,proliferation on HUVEC and A549 cell,the effect of Gambo-genic Acid(GNA)on cancer angiogenesis wasd iscussed.Methods Cell viability was determined by MTT assay.The colony forming efficiency rates of HUVEC cells detected by plate colony formation assay.The anti-angiogenic effect of GNA was examined by tube for-mation assay.Wound healing assay and transwell assay were employed to test the effect of GNA on HUVEC migration.The protein ex-pression of VEGF and HIF-1αin cells were measured by Western-blotting.Results The MTT assay showed that the viability of HU-VEC cells decreased as the concentrations of GNA.Our data demonstrated that GNA inhibited HUVEC proliferation by plate colony for-mation assay.GNA inhibited HUVEC tube formation and migration.GNA inhibited the expression of VEGF and HIF-1α.Conclusion GNA inhibits angiogenesis by down-regulation of VEGF and HIF-1α.