[Detection of stable expression of human interlukin-2 gene in transfected keratinocytes].
OBJECTIVE
To investigate the stable expression and secretion of human interlukin-2 gene in transfected keratinocytes.


METHODS
Keratinocytes were transfected with lipofectamine and selected by G418. Then the samples were analyzed with the techniques of DNA dot blot, RNA dot blot, hybridization in situ, immunohistochemistry, Western blot and MTT.


RESULTS
The positive signals were observed in transfected keratinocytes by DNA dot blot, RNA dot blot, hybridization in situ and immunohistochemistry. With Western blot analysis, a specific band exhibiting a molecular weight of 15,000 was detected in transfected keratinocytes, which was in acordance with that of IL-2. The expression of IL-2 can maintain for up to 1 month. The amounts of IL-2 in the supernatants of two and four passages transfected keratinocytes were 27.7 U/ml and 15.0 U/ml, respectively.


CONCLUSIONS
Keratinocytes have the potential for stable gene expression and secretion of active transgene products. Thus, it is possible to use keratinocytes as a target cell for gene transfection, gene expression and even gene therapy.
Liao
W
W
Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032.
Liu
Y
Y
Ye
L
L
D002478:Cells, Cultured; D015870:Gene Expression; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D014162:Transfection; D014170:Transformation, Genetic
D007376:Interleukin-2