POFUT1对人胃腺癌裸鼠移植瘤微血管生成的抑制作用

目的探讨POFUT1对人胃腺癌裸鼠移植瘤微血管生成的影响及其作用机制。方法制备人胃腺癌BGC823的POFUT1慢病毒低表达及对照组、过表达及对照组细胞模型，分别命名为shRNA-POFUT1-BGC82组、shRNA control-BGC823组、PLV-POFUT1-BGC823组及PLV control-BGC823组。采用皮下成瘤的方法，分别用这4组细胞构建裸鼠皮下移植瘤模型（每组5只），观察和测算肿瘤体积。30d后处死裸鼠并取瘤，采用免疫组化方法检测裸鼠瘤组织中VEGF和CD31的表达；Western印迹法检测裸鼠瘤组织中POFUT1和VEGF的表达。结果20只裸鼠全部成瘤，实验过程无死亡。与PLV control组相比，PLV-POFUT1组裸鼠肿瘤体积显著增加（P<0.05）；shRNA-POFUT1较shRNA control组裸鼠肿瘤体积明显偏小（P<0.05）。免疫组化结果显示，与PLV contro1组相比，PLV-POFUT1组瘤组织中VEGF和CD31表达显著增高（P<0.05）；与shRNA control组相比，shRNA-POFUT1组则明显降低(P<0.05)。与shRNA control组相比，shRNA-POFUT1组POFUT1和VEGF蛋白表达水平均显著下调；PLV-POFUT1组较PLV contro1组，其POFUT1和VEGF蛋白水平均显著增强（P<0.05）。结论过表达POFUT1可以显著促进裸鼠移植瘤的增殖和肿瘤微血管形成，低表达POFUT1可抑制裸鼠移植瘤的增殖和肿瘤微血管形成，表明POFUT1在人胃 腺癌裸鼠移植瘤的微血管形成中起着重要的作用，其机制可能与VEGF密切相关。
ObjectiveTo investigate the effect of POFUT1 on angiogenesis of human gastric adenocarcinoma xenografts in nude mice. MethodsHuman gastric adenocarcinoma BGC823 cells were transfected with shRNA-POFUT1, shRNA control, PLV-POFUT1 and PLV contro1, respectively. The transfected BGC823 cells were inoculated in nude mice. The volume of transplanted tumors was measured and the tumor growth curve was generated. The tumor-bearing nude mice were sacrificed and the tumors were harvested 30 days after transplantation. The expression of VEGF and CD31 was determined by immunohistochemistry and the expression of POFUT1 and VEGF protein was detected by Western blotting. ResultsTwenty tumor-bearing nude mice were in good condition during the experiments. Compared with PLV control group, the volume of tumor in PLV-POFUT1 group was increased significantly(P<0.05); whereas the tumor volume in shRNA-POFUT1 group was significantly lower than that in shRNA control group(P<0.05). Immunohistochemistry and Western blotting showed that the expression levels of VEGF and CD31 in PLV-POFUT1 group were significantly higher than those in control group (P<0.05), while the expression levels of POFUT1, VEGF and CD31 in the ShPOFUT1 group were significantly lower than those in control group (P<0.05). ConclusionOver-expression of POFUT1 significantly promotes the proliferation and angiogenesis of transplanted human gastric edunocarcinoma in nude mice, while low expression of POFUT1 can inhibit the proliferation and tumor angiogenesis, which indicates that POFUT1 may play an important role in the angiogenesis of human gastric adenocarcinoma xenografts, which may be closely related to VEGF