肿瘤坏死因子α干预小鼠成骨细胞cbfa1/runx2基因的表达

Tumor necrosis factor-alpha influences cbfa1/runx2 gene expression in mouse osteoblasts

背景:肿瘤坏死因子α可降低牙周膜纤维细胞碱性磷酸酶的活性,抑制牙周膜纤维细胞向成骨细胞的功能转化。目的:观察肿瘤坏死因子α对小鼠成骨细胞生长及cbfa1/runx2基因表达的影响。方法:取生长良好的小鼠成骨细胞系MC3T3/E1细胞,分别以20,40,60,80μg/L的肿瘤坏死因子α进行干预,以正常培养的细胞作为对照。采用RT-PCR法检测MC3T3/E1细胞cbfa 1/runx2mRNA的表达;PNPP法测定碱性磷酸酶活性;MTT法检测细胞活力。结果与结论:正常培养的MC3T3/E1细胞cbfa1/runx2 mRNA呈阳性表达,随着肿瘤坏死因子α浓度的增高,其表达水平逐渐下降。同时MC3T3/E1细胞活力和碱性磷酸酶活性也随肿瘤坏死因子α浓度的增高而下降。提示肿瘤坏死因子α可抑制MC3T3/E1细胞生长,而cbfa1/runx2可能参与了成骨细胞的分化过程。

BACKGROUND：Tumor necrosis factor-alpha （TNF-α） can decrease alkaline phosphatase （ALP） activity in periodontal ligament fibroblasts and inhibit the functional transformation between periodontal ligament fibroblasts and osteoblasts. OBJECTIVE：To investigate the effects of TNF-α on the growth of mouse osteoblasts and cbfa1/runx2 gene expression. METHODS：Well growing mouse osteoblast line MC3T3/E1 were interfered with 20, 40, 60, 80 μg/L TNF-α. The normally cultured cells served as controls. cbfa1/runx2 mRNA expression in osteoblast line MC3T3/E1 was detected by RT-PCR. ALP activity was determined by PNPP method and cell viability was measured by MTT assay. RESULTS AND CONCLUSION：cbfa1/runx2 mRNA expression was observed in the normally cultured osteoblast line MC3T3/E1. With increasing concentration of TNF-α concentration, cbfa1/runx2 mRNA expression, MC3T3/E1 cell viability and ALP activity were gradually decreased. Results suggested that TNF-α can inhibit osteoblast line MC3T3/E1 growth and cbfa1/runx2 may be involved in osteoblast differentiation.