中国组织工程研究2012，Vol.16Issue(19)：3421-3426,6.DOI:10.3969/j.issn.1673-8225.2012.19.001
沉默膜联蛋白A1基因影响兔骨髓间充质干细胞的增殖及成骨分化能力
Effects of silencing annexin A1 gene on proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND: Annexin A1 (ANXA1) protein is involved in regulation of cell proliferation and differentiation. Steroid-induced osteonecrosis of femoral head is related to altered proliferation and osteogeneic differentiation of bone marrow mesenchymal stem cells (BMSCs).OBJECTIVE: To investigate the effects of silencing ANXA1 gene on proliferation and osteogenic differentiation of rabbit BMSCs. METHODS: New Zealand white rabbit BMSCs were cultured in vitro and divided into three groups. In the RNA interference group, BMSCs were transfected with lentiviral vectors carrying short hairpin RNA (LV-shANXA1) to down regulate the expression of ANXA1 gene. In the negative control group, BMSCs were transfected with lentiviral vectors carrying nonsense gene sequences (LV-shANXA1-NC). In the blank control group, there was no intervention.RESULTS AND CONCLUSION: The positive rate of CD44, CD105 and CD45 in BMSCs after purification was 97.2%, 95.8 % and 2.5%, respectively. The infection efficiency of lentivirus was 80% at multiplicity of infection (MOI) of 50 and 95% at MOI of 100 in BMSCs after selection by puromycin. Real-time PCR and western blot results showed that the silencing efficiency was above 72.4% at MOI of 50. After down regulating the expression of ANXA1 gene, significant growth inhibition could be found in BMSCs; the alkaline phosphatase positive rate and alkaline phosphatase activity decreased, and the alizarin red staining results showed negative reaction in the subsequent induction of osteogenic differentiation in the cells. The experimental results show that silencing ANXA1 gene reduces the proliferation and osteogenic differentiation capacity of BMSCs, which may be one of mechanisms underlying steroid-induced avascular necrosis of femoral head.关键词
膜联蛋白A1/基因沉默/骨髓间充质干细胞/成骨分化/细胞增殖/激素性股骨头缺血性坏死分类
医药卫生引用本文复制引用
梁博伟,潘新元,赵劲民,殷国前,胡峰..沉默膜联蛋白A1基因影响兔骨髓间充质干细胞的增殖及成骨分化能力[J].中国组织工程研究,2012,16(19):3421-3426,6.基金项目
国家自然科学基金资助项目(30860285),课题名称:早期激素性股骨头缺血性坏死的蛋白质组学动态分析. （30860285）
