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ABSTRACT: To investigate the effects of wild type p16 gene on the proliferation and metabolism of human keloid fibroblasts.
Eukaryotic expression vector pcDNA3-p16 was constructed and imported into KFb by gene transfection mediated by liposome. And the positive clones were screened by G418. The transfected and untransfected KFbs were stained by Immunocytochemical method. The expression of p16 protein was observed. The changes of the proliferation and DNA synthesis of KFb before and after transfection were observed and compared by drafting cell growth curve and by (3)H-TdR incorporation method.
The recombinant vector pcDNA3-p16 was successfully constructed and identified by enzyme digestion. The positive clones were identified by G418 selection for 10 days from transfected KFb and with p16 protein expression. The growth rate of transfected KFb slowed down obviously and its DNA synthesis decreased significantly (P < 0.05) when compared with those of normal KFb.
p16 gene might inhibit the growth and DNA synthesis of KFb.
Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns 08/2003; 19(4):226-8.