红花组分HSYA对人胃腺癌BGC-823移植瘤裸鼠VEGF蛋白、KDR与缺氧诱导因子表达的影响

Effects of HSYA on protein and mRNA expression of KDR,HIF-1α and protein expression of VEGF in nude mice with BGC-823 transplantation tumor

目的:研究红花组分羟基红花黄色素A(HSYA)对人胃腺癌皮下移植瘤裸鼠血管内皮生长因子(VEGF)蛋白、含激酶插入区受体(KDR)和缺氧诱导因子(HIF-1α)mRNA与蛋白表达的影响。方法:采用BALB/C nu/nu裸小鼠接种人胃腺癌细胞株BGC-823于右前肢腋部皮下建立裸鼠人癌移植瘤模型,随机分为模型组、对照组、HSYA高、低剂量组(0.056g/L、0.028g/L),观察抑瘤作用,酶联免疫吸附测定(ELISA)法检测瘤组织VEGF与HIF-1α蛋白以及血清VEGF蛋白表达;蛋白印迹(Western blotting)法测定瘤组织KDR磷酸化蛋白的表达;实时荧光定量PCR(RTFQ-PCR)法检测瘤组织KDR及HIF-1αmRNA表达。结果:HSYA低剂量组抑瘤明显,该组瘤组织及血清VEGF蛋白表达降低,瘤组织KDR磷酸化蛋白表达减弱,与模型组比较差异明显(P<0.05,P<0.01);HIF-1α蛋白表达较模型组瘤组织明显减少(P<0.01)。KDR mRNA表达与模型组相比减弱,差异有统计学意义(P<0.05)。结论:一定浓度的HSYA抑制肿瘤生长的可能机制与抑制VEGF、HIF-1α蛋白的表达,减弱KDR蛋白磷酸化及其基因表达,从而抑制内皮细胞活化阻碍肿瘤血管新生以及降低肿瘤缺氧微环境对血管生成的诱导有关。

Objective： To investigate the effects of Hydroxy Safflor yellow A（HSYA） on the protein and mRNA expression of KDR, HIF-1 α and protein expression of VEGF in serum or transplantation tumor of gastric adenocarcinoma cell line BGC-823 in nude mice. Methods： The BGC-823 cells was subcutaneouly injected into the right anterior arrnpit of BALB/C nu/nu nude mice and established the animal model of transplantation tumor. The experimental groups were treated with HSYA at concentration of 0.056g/L and 0.028g/L and cyclophosphamide at 2g/L, or with physiologic saline. The tumor inhibitory effect was observed, and the protein expression of transplantation tumor＇s VEGF, HIF-1 a and serum VEGF were detected by enzyme linked immunosorbent assay, and the KDR protein expression was detected by Western blotting. The mRNA expression of KDR and HIF-1 et in transplantation tumor were detected by real time-fluorescent quantitation PCR respectively. Results： The IR in the group with HSYA at the concentration of 0.028g/L is higher than in the group with normal sodium. The protein expression of VEGF, phosphorylated KDR and HIF-1 α has significant difference after treatment with HSYA at the concentration of 0.028g/L as compared with physiologic saline-treated group（P〈0.05, P〈0.01）. The mRNA expression of KDR in 0.028g/L HSYA group is less obviously than in the physiologic saline.-treated group（P〈0.05）. Conclusion： The possible mechanism of＇HSYA in given concentration to antagonize tumor angiogenesis may be related with inhibiting the protein expression of VEGF, and HIF-1 α and weakening the phosphorylation of KDR protein and its gene expression to inhibit the activation of endotheliocyte and impede the inducion of tumor oxygen-poor microenvironment to angiogenesis.