成人成骨细胞和MG-63细胞株孕激素受体的表达

Expression of progesterone receptors in human normal osteoblasts and MG-63 cells

目的 观察成人成骨细胞和MG-63细胞株孕激素受体(progesterone receptor,PR)亚型表达的情况。方法用改良胶原酶消化法从正常成人松质骨中分离成骨细胞,观察细胞形态,钙钴法行碱性磷酸酶染色,Van Gieson法行Ⅰ型胶原染色,茜素红行矿化结节染色;半定量RT-PCR检测骨钙素和PR亚型mRNA表达;Western blot测定PR蛋白质表达。结果所分离培养的细胞具有成骨细胞的形态特征,保持了其在体内的功能;人成骨细胞和MG-63细胞株均表达PRA、PRB mRNA和蛋白质。结论人成骨细胞和MG-63细胞株可能受孕激素的影响。

Objective To observe the expression of progesterone receptors in human osteoblasts and human osteosarcoma cell line MG-63. Methods The human osteoblasts were isolated from the trabecular bone by collagenase digestion. The expression of alkaline phosphatase, collagen type Ⅰ, mineralized nodes and osteocalcin were studied with modified Gonori's assay, van Gieson's staining, alizarin red-S staining and RT-PCR, separately. The expression of PRA and PRB mRNA and protein was measured by RT-PCR and Western blot in human osteoblasts and MG-63 cells. Results The isolated normal human osteoblasts possessed the phenotypes and functions of osteoblasts in vivo. PRA and PRB mRNA and protein were espressed in both human osteoblasts and MG-63 cells. Conclusions Human osteoblasts and MG-63 cells are the target cells of progesterone through its receptors.