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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。
Objective To investigate the possible molecular mechanism of Hedgehog signaling pathway regulating in vitro vascularization of human gastric cancer cell line SGC-7901. Methods SGC-7901 cells and human umbilical vein endothelial cells (HUVEC) were cultured routinely. SGC-7901 cells were divided into control group A (untreated group), experimental group B (final target concentration of 5 μmol / L and experimental group C with a final target concentration of 10μmol / L. After cultured for 48 hours, the expression of Glioma-associated oncogene homolog (Gli1) and platelet-derived growth factor- The expression of PDGF-D in gastric cancer SGC-7901 cells was detected by flow cytometry. The changes of vascular formation ability before and after treatment were detected by tubule formation test and vascular mimicry test. The expression of PDGF-D was detected by RT-PCR and Western blot Gli1, PDGF-D mRNA and protein expression changes, and the two mRNA and protein expression levels were analyzed. Results The expressions of Gli1 and PDGF-D in SGC-7901 cells were significantly higher than those in the control group (P <0.05). The SGC-7901 cells in vitro were decreased in vitro and in a dose-dependent manner (P <0.05) . The mRNA and protein expressions of Gli1 and PDGF-D in the experimental group were significantly inhibited compared with the control group (P <0.05), and the mRNA and protein expressions of Gli1 and PDGF-D in the experimental group were positively correlated (r = 0.829, r = , P <0.05). Conclusion The Hedgehog signaling pathway may regulate the expression of PDGF-D through Gli1 and further promote tumor angiogenesis.