Senescence of human keloid fibroblasts induced by wild p16 gene in vitro / 中华创伤杂志

ABSTRACT

Objective To investigate the biological effect of wild p16 gene on senescence of human keloid fibroblasts (HKFb) and discuss the feasibility of gene therapy on keloid. Methods Firstly, the HKFb was transfected with pcDNA3-p16 and the positive clones selected by G418. Then, the fibroblast changes were investigated by determining growth curve, cycle profile and ultrastructure of the transfected HKFb. Meanwhile, morphological changes of the transfected HKFb were identified by senescence associated-?-galactosidase staining. Results After HKFb was transfected by p16, the morphology of HKFb changed obviously from the third passage on. The cell proliferation was inhibited markedly and the cell cycle profile analysis showed that about 96.7% cells were trapped in stage G1. The electroscopic analysis of HKFb also showed that the lysosomes were overloaded. Senescence associated-?-galactosidase staining identified the senescence of HKFb. Conclusions The wild p16 gene can induce the HKFb into senescence in advance, as may provide experimental basis for gene therapy on human keloid.