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目的：观察含外源性人碱性成纤维细胞生长因子（ｈｂＦＧＦ）基因的角朊细胞（ｈｕｍａｎｋｅｒａｔｉｎｏｃｙｔｅ，ＨＫ），对真皮成纤维细胞（ｈｕｍａｎｄｅｒｍａｌｆｉｂｒｏｂｌａｓｔ，ＨＤＦｉｂ）增殖的影响；探讨转ｈｂＦＧＦ基因ＨＫ对创面愈合的生物学效应及作用机理，为这种转基因ＨＫ细胞用于创面基因治疗的可行性提供实验依据．方法：收集含真核表达质粒ｐｃＤＮＡ３－ｈｂＦＧＦ的ＨＫ细胞培养上清，加入培养的ＨＤＦｉｂ细胞内；ＭＴＴ法测定细胞的增殖率、３Ｈ－ＴｄＲ掺入法测定细胞ＤＮＡ合成率、流式细胞仪分析细胞增殖周期．结果：加入转基因ＨＫ细胞培养上清的ＨＤＦｉｂ细胞，培养７２ｈ后，较正常对照组、加正常ＨＫ培养上清组、加转染空载体ｐｃＤＮＡ３－ｎｅｏ的ＨＫ培养上清组，其细胞增殖分别增加１．０，１．５，３．０倍（Ｐ＜０．０１）；细胞ＤＮＡ合成率分别提高１．６，２．３，３．４倍（Ｐ＜０．０１）；ＨＤＦｉｂ细胞周期分析显示加入转基因ＨＫ细胞培养上清２４ｈ时细胞处于Ｓ期的比例较其余对照组增加明显，至７２ｈ时无差别．结论：转染ｈｂＦＧＦ基因真核表达质粒的ＨＫ细胞可以分泌活性物质促进ＨＤＦｉｂ细胞ＤＮＡ合成及增殖；机理为刺激ＨＤＦｉｂ进?
OBJECTIVE: To observe the effect of exogenous human basic fibroblast growth factor (hbFGF) on the proliferation of human dermal fibroblast (HDFib) and to investigate the effect of hbFGF gene HK on the wound surface The biological effects of healing and mechanism of action, provide experimental evidence for the feasibility of genetically modified wounds in this genetically modified HK cells. Methods: The supernatant of HK cells containing the eukaryotic expression plasmid pcDNA3-hbFGF was collected and cultured in HDFib cells. The cell proliferation rate was measured by MTT assay. The DNA synthesis rate was determined by 3H-TdR incorporation assay. Flow cytometry Cell cycle. Results: After cultured for 72h, HDFib cells cultured in the supernatant of transgenic HK cells were added with normal HK group and HK group with empty vector pcDNA3-neo 1.0, 1.5 and 3.0 folds (P <0.01). The rate of DNA synthesis increased by 1.6, 2.3 and 3.4 times (P <0.01) respectively. The cell cycle analysis The results showed that the proportion of cells in S phase increased 24 h after transfection with HK cell culture supernatant, and no significant difference was observed at 72 h. CONCLUSION: HK cells transfected with eukaryotic expression plasmid of hbFGF gene can secrete active substances to promote DNA synthesis and proliferation of HDFib cells.