关键词：转化生长因子β；骨形成蛋白-2；基因转染；细胞分化
摘要 目的: 探讨骨形成蛋白-2(BMP-2)和转化生长因子β(TGFβ)对细胞增殖分化的调控作用及其机制。方法: 构建含有人BMP-2基因的噬菌粒表达载体pBK-B2,利用脂质体转染方法将其导入NIH3T3细胞,通过G-418筛选获得阳性细胞克隆。细胞原位杂交、免疫组化法检测BMP-2基因的稳定转染及表达,并检测了外源性TGFβ对转染细胞增殖活力及碱性磷酸酶活性和骨钙素含量的影响。结果: TGFβ(50 ng/ml)促进BMP-2转染细胞的增殖,但是抑制其碱性磷酸酶活性和骨钙素的含量。结论: TGFβ的生物学效应与多种因素密切相关,BMP和TGF-β在细胞生长分化过程中具有协同作用。
Effects of TGFβ on BMP-2 Gene Transfection Cellsin Vitro
Si Xiaohui, Yang Lianjia, Jin Yan
Department of Oral Pathology, Stomatological College, the Fourth Military MedicalUniversity
Abstract
Objective： To elucidate the mechanism of bone morphogenetic protein-2(BMP-2)and transforming growth factor β(TGFβ) on cell differentiation. Methods: A phagemidexpression vector for human BMP-2 (pBK-B2) was constructed and transfected into the NIH3T3cells by using lipofectamine. Positive cell clones were selected with G418. The stabletransfection and expression of BMP-2 were determined by using in situ hybridization andimmunohistochemistry methods. The proliferativity of the transfected cells treated withTGFβ were assayed by MTT method. Alkaline phosphatase(ALP) and osteocalcin (OC) production were also measured. Results: Theproliferation of the transfected cells was stimulated after being treated with TGFβ (50 ng/ml). But the ALP activity and OC production wereinhibited. Conclusion: The effects of TGFβ are closely related to many factors, and BMP and TGFβ may be synergistic in cell proliferation and differentiation.
Key words: transforming growth factor β bonemorphogenetic protein 2 gene transfection cell differentiation
骨形成蛋白(bone morphogenetic proteins,BMPs)作为骨的生长因子属于TGFβ超家族的一个成员 ......
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