Hedgehog信号通路对人胃癌SGC-7901细胞体外血管形成的调控

目的探讨Hedgehog信号通路调控人胃癌SGC-7901细胞体外血管形成的可能分子机制。方法常规培养SGC-7901细胞和人脐静脉内皮细胞(Human umbilical vein endothelial cell,HUVEC),将SGC-7901细胞分为对照组A(未给药)、实验组B(环靶明终浓度5μmol/L)及实验组C(环靶明终浓度10μmol/L),培养48 h后,免疫荧光检测各组胶质瘤相关癌基因1(Glioma-associated oncogene homolog,Gli1)和血小板源性生长因子-D(Platelet-derivedgrowth factor D,PDGF-D)在胃癌SGC-7901细胞中的表达;通过小管形成试验和血管拟态试验检测细胞处理前后血管形成能力的变化;RT-PCR和Western blot检测细胞处理前后Gli1、PDGF-D的mRNA和蛋白表达变化,并对二者mRNA和蛋白表达水平进行相关性分析。结果 Gli1和PDGF-D在各组SGC-7901细胞中均有表达;实验组SGC-7901细胞的体外血管形成能力较对照组下降,且呈剂量依赖性,差异具有统计学意义(P<0.05);实验组Gli1和PDGF-D的mRNA和蛋白表达较对照组均受到显著抑制,差异有统计学意义(P<0.05),且二者mRNA和蛋白表达水平呈正相关(r=0.829,r=0.786,P<0.05)。结论 Hedgehog信号通路可能通过Gli1来调控PDGF-D的表达,进而促进肿瘤血管的形成。

Role of Hedgehog signaling pathway in angiopoiesis in vitro of human gastric cancer SGC-7901 cells

Objective To investigate the role of Hedgehog(Hh) signaling pathway in angiopoiesis of human gastric cancer SGC-7901 cells in vitro as well as the relevant mechanism.Methods SGC-7901 and Human umbilical vein endothelial cell(HUVEC) were cultured by routine method.SGC-7901 cells were divided into control group A and test groups B and C.The cells in group A were untreated,while those in groups B and C with cyclopamine at final concentrations of 5 and 10 μmol / L respectively,and further cultured for 48 h.The expressions of glioma-associated oncogene homolog(Gli1) and platelet-derived growth factor D(PDGF-D) in SGC-7901 cells of various groups were determined by IFA,while the ability of angiopoiesis before and after treatment by tube formation assay and vasculogenic mimicry(VM) formation assay.The expressions of Gli1 and PDGF-D at mRNA and protein levels were determined by RT-PCR and Western blot respectively,between which the relationship was analyzed.Results Gli1 and PDGF-D were expressed in SGC-7901 cells of various groups.As compared with those in control group,the abilities of angiopoiesis in vitro of SGC-7901 cells in test groups showed a dose-dependent decrease(P < 0.05),while the expressions of Gli1 and PDGF-D at mRNA and protein levels decreased significantly(P < 0.05).However,the expressions of Gli1 at both mRNA and protein levels were positively related to those of PDGF-D,with r values of 0.829 and 0.786 respectively(P < 0.05).Conclusion Hh signaling pathway may promote the angiopoiesis of tumor by regulating the expression of PDGF-D with Gli1.