Role of p21~(wafl)in the proliferation and differentiation of the human osteosarcoma cell line(HOS-8603) regulated by 1,25(OH)_2D_3 / 中国病理生理杂志

ABSTRACT

AIM: To explore the possible role of p21 wafl gene in the effect of 1,25-dihydroxyvitamin D 3 on a human osteosarcoma cell line (HOS-8603) proliferation and differentiation. METHODS: The effect of 1,25(OH) 2D 3 on the expression of p21 wafl mRNA was determined by quantitative reverse transcription-polymerase chain reaction. The human osteosarcoma cell line stably expressing the p21 wafl mRNA, which named HOS-p21 wafl , was constructed by lipofectamine transfection, and the expression of p21 wafl protein was detected by Western blot. Then the growth curve of the HOS-p21 wafl cells and the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells were investigated. RESULTS: The expression of endogenous p21 wafl mRNA in HOS-8603 cells was induced by 1,25(OH) 2D 3 in a time-dependent manner, reaching the maximum at 4 h, and remaining the inducible action for up to 24 h over basal levels. The HOS-p21 wafl cells grew much slower than the control cells, only reaching 50% of the control ones when cultured for up to 6 days. Histochemical analysis showed that the expression of alkaline phosphatase (AKP) in the HOS-p21 wafl cells increased remarkably.CONCLUSION: These results suggest that the p21 wafl mRNA expression induced by 1,25(OH) 2D 3 is one of important mechanisms responsible for the cellular effects of the hormone on HOS-8603 cells.