Lafage-Proust M H, Wesolowski G, Ernst M, Rodan G A, Rodan S B
Department of Bone Biology and Osteoporosis Research, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
J Cell Physiol. 1999 Jun;179(3):267-75. doi: 10.1002/(SICI)1097-4652(199906)179:3<267::AID-JCP4>3.0.CO;2-0.
Clonal cell lines were established from adult rat tibia cells immortalized with SV-40 large T antigen. One clone (TRAB-11), in which retinoic acid (RA) induced alkaline phosphatase (AP) activity, was selected for further study. The TRAB-11 cells express high levels of type I collagen mRNA, type IV collagen, fibronectin, practically no type III collagen, little osteopontin, and no osteocalcin. RA stimulates proliferation of TRAB-11 cells (starting at 10 pM) and survival (starting at 100 pM). TRAB-11 cells synthesize fibroblast growth factor-2 (FGF-2), which has potent autocrine mitogenic effects on these cells and acts synergistically with RA. TRAB-11 cells attach better to type IV collagen than to fibronectin or laminin. Cell attachment to type IV collagen is increased by RA and decreased (65%) by an antibody directed against alpha1beta1 integrin. RA up-regulates steady-state levels of alpha1, mRNA without affecting beta1 mRNA expression. In conclusion, we report the establishment of a clonal cell line from the outgrowth of adult rat tibiae which is highly sensitive to RA in its growth and survival in culture, apparently as a result of integrin-mediated cell interaction with extracellular matrix proteins.
从用SV - 40大T抗原永生化的成年大鼠胫骨细胞建立了克隆细胞系。选择了一个克隆（TRAB - 11）进行进一步研究，在该克隆中视黄酸（RA）可诱导碱性磷酸酶（AP）活性。TRAB - 11细胞高水平表达I型胶原mRNA、IV型胶原、纤连蛋白，几乎不表达III型胶原，骨桥蛋白表达很少，且不表达骨钙素。RA刺激TRAB - 11细胞增殖（从10 pM开始）和存活（从100 pM开始）。TRAB - 11细胞合成成纤维细胞生长因子 - 2（FGF - 2），其对这些细胞具有强大的自分泌促有丝分裂作用，并与RA协同发挥作用。TRAB - 11细胞与IV型胶原的附着比与纤连蛋白或层粘连蛋白更好。RA可增加细胞与IV型胶原的附着，而抗α1β1整合素抗体可使其减少（65%）。RA上调α1 mRNA的稳态水平，而不影响β1 mRNA的表达。总之，我们报道了从成年大鼠胫骨长出的细胞建立的一个克隆细胞系，该细胞系在培养中的生长和存活对RA高度敏感，这显然是整合素介导的细胞与细胞外基质蛋白相互作用的结果。
