Hey1基因过表达对小鼠单核细胞RAW264.7向破骨细胞分化的影响

The effect of hey1 overexpression on osteoclastogenesis capacity of murine monocytes RAW264.7

目的:观察Hey1基因过表达对小鼠单核细胞RAW264.7向破骨细胞分化的影响。方法:用Lipofectamine 2000将Hey1表达质粒转染RAW264.7细胞系;10μg/mL Blasticidin筛选稳定转染的细胞克隆,RT-PCR检测Hey1基因表达水平;用RANKL诱导RAW264.7细胞向破骨细胞分化,抗酒石酸磷酸酶(TRAP)染色后进行破骨细胞计数。结果:Blasticidin筛选出的细胞株Hey1 mRNA表达水平显著高于正常RAW264.7细胞;在50 ng/mL RANKL诱导条件下,Hey1过表达的RAW264.7细胞分化形成的破骨细胞数显著低于正常RAW264.7细胞(P<0.05)。结论:Hey1具有抑制小鼠单核细胞RAW264.7向破骨细胞分化的作用。

AIM ： To observe the effect of Heyl overexpression on the differentiation of murine monocytes of RAW264.7 into osteoelasts. METHODS ： Expression plasmid carrying Hey1 mRNA encoding sequence was transfected into RAW264.7 cells by lipofectamine 2000. Stably transfected cells were screened by blasticidin at concentration of 10μg/mL. Heyl mRNA expression was analyzed by RT-PCR. RANKL was used to induce RAW264.7 cells to differentiate into osteoelasts. The osteoclasts were counted after tartrate-resistant acid phosphatase （TRAP） staining. RE- SULTS ： Hey1 mRNA expression level of the cell line screened by blastieidin was significantly higher than that of parental RAW264.7 Cells. In the presence of 50ng/ml RANKL, more osteoclasts were doserved in the Heyl over expressed RAW 264.7 cells than in the parental RAW264.7 cell （ P 〈 0.05 ）. CONCLUSION ： Hey1 has an inhibitory effect on the differentiation of murine RAW264.7 monocytes into osteoclasts.