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目的 探讨重组骨形态发生蛋白-7腺病毒载体(AdCMV-BMP7)在人真皮成纤维细胞获得稳定表达的可行性。方法重组腺病毒载体AdCMV-BMP7经293细胞包装、扩增后,转染人真皮成纤维细胞。转染48 h后以免疫组织化学、RT-PCR等方法检测hBMP7基因在人真皮成纤维细胞内的表达情况。结果 在合适的滴度下,重组腺病毒载体AdCMV-BMP7对成纤维细胞的增殖能力无明显影响。转染48 h后免疫细胞化学显示已转染AdCMV-BMP7的成纤维细胞内有大量的hBMP7表达,RT-PCR也检测到特异的hBMP7基因片段的表达。结论 重组腺病毒载体AdCMV-BMP7能够被转导入成纤维细胞内,并稳定表达hBMP7。
Objective To investigate the feasibility of recombinant human bone morphogenetic protein-7 adenovirus vector (AdCMV-BMP7) for stable expression in human dermal fibroblasts. Methods Recombinant adenovirus AdCMV-BMP7 was packaged by 293 cells and amplified. Human dermal fibroblasts were transfected. Forty-eight hours after transfection, the expression of hBMP7 gene in human dermal fibroblasts was detected by immunohistochemistry and RT-PCR. Results Under suitable titers, recombinant adenovirus vector AdCMV-BMP7 had no significant effect on the proliferation of fibroblasts. Immunocytochemistry 48 h after transfection showed that a large amount of hBMP7 was expressed in the fibroblasts transfected with AdCMV-BMP7, and a specific hBMP7 gene fragment was also detected by RT-PCR. Conclusion The recombinant adenovirus vector AdCMV-BMP7 can be transduced into fibroblasts and stably express hBMP7.