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目的观察结缔组织生长因子(CTCF)和多聚腺苷酸结合蛋白相互作用蛋白1(PAIP-1) mRNA在MG-63细胞培养不同阶段的表达,研究17β-雌二醇(E_2)对MG_63细胞CTGF和PAIP-1 mRNA表达的影响。方法半定量RT-PCR检测MC-63细胞碱性磷酸酶(ALP)、骨钙素(OC)和Ⅰ型胶原mRNA的表达,半定量RT-PCR和Northern印迹方法检测MG-63细胞CTGF和PAIP-1 mRNA表达,改良的钙-钴法ALP染色、van Gieson法Ⅰ型胶原染色和0.1%茜素红矿化结节染色。结果(1)半定量RT-PCR和组织化学染色结果均证实,MG-63细胞接种培养后0～9d为细胞增殖阶段,7～18d为基质成熟阶段,17～18d后为基质矿化阶段；(2)在MG-63细胞培养的不同阶段,均检测到CTGF和PAIP-1 mRNA的表达,E_2可显著下调MG_63细胞CTGF mRNA的表达,并呈剂量依赖关系(P＜0.01),但时间依赖性关系不明显。17β-E_2对MG-63细胞PAIP-1 mRNA表达无明显作用(P＞0.05)。结论E_2可剂量依赖性下调MG-63细胞CTGF mRNA的表达,但对PAIP-1 mRNA表达无明显作用。
Objective To investigate the expression of connective tissue growth factor (CTCF) and polyclonal-binding protein-1 (PAIP-1) mRNA in different stages of MG-63 cell culture and to investigate the effect of 17β-estradiol CTGF and PAIP-1 mRNA expression. Methods The expression of alkaline phosphatase (ALP), osteocalcin (OC) and collagen Ⅰ mRNA in MC-63 cells were detected by semi-quantitative RT-PCR. CTGF and PAIP were detected by semi-quantitative RT-PCR and Northern blotting -1 mRNA expression, modified calcium-cobalt ALP staining, van Gieson method type I collagen staining and 0.1% alizarin red mineralized nodule staining. Results (1) The results of semi-quantitative RT-PCR and histochemical staining confirmed that MG-63 cells were in cell proliferation stage 0-9 days after inoculation culture, maturation phase in 7-18 days and matrix mineralization stage in 17-18 days. (2) The expressions of CTGF and PAIP-1 mRNA were detected in different stages of MG-63 cell culture. E_2 could down-regulate the expression of CTGF mRNA in MG_63 cells in a dose-dependent manner (P <0.01) Sexual relationship is not obvious. 17β-E 2 had no effect on PAIP-1 mRNA expression in MG-63 cells (P> 0.05). Conclusion E_2 can down-regulate the expression of CTGF mRNA in MG-63 cells in a dose-dependent manner, but it has no effect on the expression of PAIP-1 mRNA.