p21^(wafl)在1,25(OH)_2D_3调节人骨肉瘤细胞系HOS-8603增殖分化中的作用

Role of p21^(wafl)in the proliferation and differentiation of the human osteosarcoma cell line(HOS-8603) regulated by 1,25(OH)_2D_3

目的 :探讨p2 1wafl在 1,2 5 (OH) 2 D3调节人骨肉瘤细胞系HOS - 860 3增殖中的作用。方法 :用定量RT-PCR法检测 1,2 5 (OH) 2 D3对p2 1waflmRNA表达的诱导作用 ,并构建稳定表达p2 1wafl真核表达载体的细胞系HOS -p2 1wafl进一步观察该细胞的生长及分化情况。结果 :1,2 5 (OH) 2 D3作用 2h即可诱导HOS - 860 3细胞内源性p2 1wafl的表达 ,4h达高峰 ,2 4h仍未回降。细胞过度表达p2 1wafl后生长速度明显减慢 ,培养 6d时细胞数为未转染细胞的5 0 % ;并且组织化学染色结果表明细胞的分化标志性抗原碱性磷酸酶 (AKP)的表达明显增强。结论 :表明 1,2 5(OH) 2 D3对p2

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.