BICC1对骨髓基质细胞定向分化的调控作用研究  

The role of BICC1 in osteogenic and adipogenic differentiation of bone marrow stromal cells

作　　者：杨俊英 李晓霞 YANG Jun-ying;LI Xiao-xia(Department of Pathogen Biology,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China)

机构地区：[1]天津医科大学基础医学院病原生物学系,天津300070

出　　处：《天津医科大学学报》2020年第3期199-203,共5页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助项目(81772297)。

摘　　要：目的:探讨BICC1在骨髓基质细胞向成骨细胞及脂肪细胞分化过程中的作用。方法:构建Bicc1过表达质粒Bicc1-pcDNA3.1,以pcDNA3.1为对照,分别转染小鼠骨髓基质细胞ST2。对两组细胞进行成脂和成骨诱导,在成骨诱导14 d时碱性磷酸酶染色检测成骨分化情况,在成脂诱导5 d时利用油红O染色检测脂滴形成情况;采用qRT-PCR及Western blot技术检测细胞Bicc1过表达对成骨及成脂相关因子的影响。结果:酶切及测序结果显示Bicc1过表达质粒构建成功,将其转染到ST2细胞后,Bicc1 mRNA表达水平较对照组升高15.23倍(P<0.05)。成骨诱导条件下,Bicc1过表达组ST2细胞碱性磷酸酶染色增强,成骨相关因子Osterix、碱性磷酸酶、Osteopontin、Runt相关转录因子2(Runx2)和骨钙素的mRNA和/或蛋白表达水平显著上升(均P<0.05)。成脂诱导条件下,Bicc1过表达组ST2细胞中脂滴形成减少,油红OD520较对照组明显降低(P<0.01),成脂相关因子过氧化物酶体增殖物活化受体γ(PPARγ)、CCAAT增强子结合蛋白α(C/EBPα)、脂肪酸结合蛋白(FABP4/aP2)和adipsin的mRNA及蛋白表达水平显著下降(均P<0.05)。结论:BICC1可促进骨髓基质细胞向成骨细胞分化,抑制其向脂肪细胞分化。Objective:To study the role of BICC1 in osteogenic and adipogenic differentiation of bone marrow stromal cells.Methods:Bicc1 expression construct was made and transfected into mouse marrow stromal cell line ST2.The cells transfected with the vector pcDNA3.1 served as control.The transfected cells were subjected to osteogenic treatment for 14 days or adipogenic treatment for 5 days.For the osteogenic differentiation assay,alkaline phosphatase(ALP)staining was done and the expression of osteogenic factors were examined by using qRT-PCR and Western blot.For the adipogenic differentiation assay,oil-red O staining was done and the expression of adipogenic factors were examined.Results:The identify of the Bicc1 overexpression plasmid was verified by enzyme digestion and DNA sequencing.The Bicc1 mRNA in over-expressed group was markedly increased compared with control group(15.23-fold difference,P<0.05).Following osteogenic induction,Bicc1 overexpression stimulated the differentiation of osteoblasts from ST2 cells as compared to vector transfection,as evidenced by the enhanced ALP staining and the increased mRNA and protein levels of osteogenic factors such as osterix,ALP,osteopontin,Runt-related transcription factor(Runx2)and osteocalcin(all P<0.05).Following adipogenic induction,Bicc1 overexpression inhibited the differentiation of adipocytes from ST2 cells,as evidenced by the attenuated oil-red O staining and the decreased mRNA and protein levels of adipogenic factors such as peroxisome proliferator-activated receptorγ(PPARγ),CCAAT enhancer-binding proteinα(C/EBPα),fatty acid-binding protein 4(FABP4/aP2)and adipsin(all P<0.05).Conclusion:BICC1 promotes osteogenic differentiation and suppresses adipogenic differentiation of bone marrow stromal cells.

关 键 词：Bicc1 骨髓基质细胞 分化 成骨细胞 脂肪细胞 

分 类 号：R37[医药卫生—病原生物学]