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目的 探讨人血管内皮细胞生长因子(VECF)基因修饰的免骨髓基质细胞对VEGF基因的表达及表达产物的生物学活性。方法 制备真核表达质粒pAdCMV-VEGF165,以Superfect转染试剂介导VFGF基因转染兔骨髓基质细胞,收获转染后24h至6d的转染上清和细胞,采用RT-PCR技术、免疫组化SABC法检测转染细胞中外源性VECF165基因的转录及表达,用血管内皮细胞增殖试验测定转染细胞培养上清中VEGF的生物活性。结果 VEGF基因转染的兔骨髓基质细胞可有效转录VEGF165,其分泌于培养上清中的表达产物可促进血管内皮细胞增殖,具有很强的生物活性。结论 VEGF基因转染的兔骨髓基质细胞可有效表达具有生物活性的VEGF。
Objective To investigate the expression of vascular endothelial growth factor (VEGF) gene and the biological activity of human bone marrow stromal cells modified by human vascular endothelial cell growth factor (VECF) gene. Methods The eukaryotic expression plasmid pAdCMV-VEGF165 was transfected into rabbit bone marrow stromal cells (VFGF) mediated by Superfect transfection reagent. Transfection supernatants and cells were harvested 24h to 6d after transfection. The expression of SABC Method to detect the transcription and expression of exogenous VECF165 gene in transfected cells, and to determine the biological activity of VEGF in the transfected cell culture supernatant by using the vascular endothelial cell proliferation assay. Results The VEGF gene transfected rabbit bone marrow stromal cells could effectively transcribe VEGF165, which secreted in the culture supernatant can promote the proliferation of vascular endothelial cells and have strong biological activity. Conclusion The VEGF gene transfected rabbit bone marrow stromal cells can effectively express the biological activity of VEGF.