中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (7): 439-441.

• 论著 • 上一篇    下一篇

血管内皮细胞生长因子真核表达载体在HaCaT细胞中的表达

周乃慧, 范卫新   

  1. 1. 南京医科大学第一附属医院(江苏省人民医院)皮肤 210029;
    2. 南京医科大学第一附属医院(江苏省人民医院)皮肤 210029
  • 收稿日期:2004-07-19 发布日期:2005-07-15
  • 通讯作者: 范卫新, E-mail:weixinfan@medmail.com.cn E-mail:weixinfan@medmail.com.cn
  • 基金资助:
    国家自然科学基金资助项目(3017086)

ZHOU Nai-hui, FAN Wei-xin   

  1. Department of Dermatology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2004-07-19 Published:2005-07-15

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摘要： 目的 探讨外源性人vEGF165基因在HacaT细胞中表达的可行性及其对体外培养的猪毛囊的影响。方法 通过脂质体将与增强型绿色荧光蛋白(EGFP)基因共表达的血管内皮细胞生长因子(vEGF)真核表达载体PIRES2-GFP-vEGF165。瞬时转染HaCaT细胞,应用激光共聚焦显微镜观察EGFF在HaCaT细胞内的表达,同时利用ELISA法检测vEGF在培养细胞被转染的上清液中的表达。进一步将该上清液加至体外培养的猪毛囊中,显微镜下测量毛囊的平均生长长度,并观察毛囊的形态学变化。结果 成功地将vEGF真核表达载体PIRES厂EGFP一vEGF165瞬时转染了HacaT细胞,以激光共聚焦显微镜观察可见细胞内EGFP的表达,同时用ELIsA法证实了上清液中vEGF呈高水平表达,并且该上清液可以明显促进体外培养的猪毛囊生长,延缓其进人退行期。结论 应用脂质体能够成功地将外源性人vEGF165基因转染HaCaT细胞,并进行有效表达,其表达的vEGF在体外具有促进猪毛囊生长的生物学活性。

关键词: 内皮生长因子, 增强型绿色荧光蛋白, 细胞,培养的, 毛囊

Abstract: Objective To investigate the feasibility of transient transfection of human VEGF165 gene into HaCaT cells and the effects of suPernatant from transfected cell culture on pig hair follicles in vitro.Methods PIRES2-EGFP-VEGF165 was transiently transfected into HaCaT cells with lipofectaminemTM 2000. Expression of EGFP (enhanced green fluorescence protein,EGFP) was observed by laser confocal microscopy.VEGF in the supernatant was detected by ELISA.Further,the supernatant was added to pig hair follicles cultured in vitro.The growth and morphologic changes of hair follicles were measured.Results PIRES2-EGFP-VECF165 was successfully transfected into HaCaT cells,which confirmed by laser confocal microscopy and ELISA.Moreover,the supernatant of HaCaT cells transfected with PIRES2-EGFP-VEGF165 could accelerate the growth of Pig hair follicles and Prolong their anagen phase.Conclusion PIRES2-EGFP-VEGF165 could be successfully transiently transfected into HaCaT cells,and expressed VEGF could upregulate the growth of pig hair follicles invitro.

Key words: Endothelial grwth factors, Enhanced green fluorescent protein, Hair follicle, Cells,cultured