摘要
目的:检测在8种骨肉瘤细胞系中Runx3基因启动子区域甲基化状态和Runx2基因及Runx3基因的表达情况,探讨Runx2基因和Runx3基因对骨肉瘤形成的影响。方法:采用甲基化特异PCR(MSP)对8种骨肉瘤细胞系中Runx3基因启动子区域甲基化状态进行分析,采用逆转录多聚合酶链反应(RT-PCR)检测Runx3及Runx2基因的表达情况。结果:MSP结果显示,Runx3基因启动子区域无明显过甲基化状态发生,RT-PCR结果表明,大多数骨肉瘤细胞系中Runx2基因和Runx3基因均有不同程度表达。结论:在骨肉瘤细胞系中无明显的Runx3基因特异性表达异常,Runx3基因启动子区域未见CpG岛发生广泛甲基化等表观遗传学异常改变,提示Runx3基因与骨肉瘤的发生发展无必要联系。Runx2基因虽然在成骨细胞分化、软骨细胞成熟及骨基质蛋白产生中发挥重要作用,但在骨肉瘤细胞系中均有不同程度的表达。
Objective:To study the effect of Runx3 and Runx2 on osteosarcoma, we examine the promoter methylation status of the Runx3 gene and detect the expression of the Runx3 gene and Runx2 gene in eight osteosarcoma cell lines.Methods:The methylation status of Runx3 gene was examined by methylation specific polymerase chain reaction (MSP), the expression of Runx3 and Runx2 gene were detected by reverse transcription polymerase chain reaction (RT-PCR).Results:There was not significant hypermethylation in the promoter region of the Runx3 gene.RT-PCR indicated there was a certain expression in the most osteosarcoma cell lines. Conclusion:The expression of the Runx3 gene was not significantly abnormal and the most CpG islands were not hypermethylated in the osteosarcoma cell lines, so it suggests the Runx3 gene is not correlative with the osteosarcoma.Runx2 is a specific transcription factor that can regulate organ formation, it plays a pivotal role in osteoblast differentiation and bone formation.In humans, Runx2 gene heterozygous mutation or expression insufficiency results in the clavicle or skull development disorder, but the Runx2 gene is a certain expression in osteosarcoma cell lines.