Adhesion to fibronectin stimulates proliferation of wild-type and bcr/abl-transfected murine hematopoietic cells.
Cells of most tissues require adhesion to a surface to grow. However, for hematopoietic cells, both stimulation and inhibition of proliferation by adhesion to extracellular matrix components have been described. Furthermore, it has been suggested that progenitor cells from chronic myelogenous leukemia show decreased beta1 integrin-mediated adhesion to fibronectin, resulting in increased proliferation and abnormal trafficking. However, we show here that the chronic myelogenous leukemia-specific fusion protein p210bcr/abl stimulates the expression of alpha5beta1 integrins and induces adhesion to fibronectin when expressed in the myeloid cell line 32D. Moreover, proliferation of both p210bcr/abl-transfected 32D (32Dp210) cells and untransfected 32D cells is stimulated by immobilized fibronectin. Cell cycle analysis revealed that nonadherent 32D and 32Dp210 cells are arrested in late G1 or early S phase, whereas the adherent fractions continue cycling. Although both adherent and nonadherent p210bcr/abl-transfected and parental 32D cells express equal amounts of cyclin A, a protein necessary for cell cycle progression at the G1/S boundary, cyclin A complexes immunoprecipitated from 32D cells cultured on immobilized fibronectin were found to be catalytically inactive in nonadherent but not in adherent cells. In addition, as compared with untransfected 32D cells, cyclin A immunoprecipitates from 32Dp210 cells exhibited a greatly elevated kinase activity and remained partially active irrespective of the adhesion status. The lack of cyclin A/cyclin-dependent kinase (CDK) 2 activity in nonadherent 32D cells appeared to result from increased expression and cyclin A complex formation of the CDK inhibitor p27(Kip1). Taken together, our results indicate that adhesion stimulates cell cycle progression of hematopoietic cells by down-regulation of p27(Kip1), resulting in activation of cyclin A/CDK2 complexes and subsequent transition through the G1/S adhesion checkpoint.
Krämer
A
A
III. Medizinische Klinik, Klinikum Mannheim, Universität Heidelberg, Wiesbadener Str. 7-11, 68305 Mannheim, Germany. akraemer@rumms.uni-mannheim.de
Hörner
S
S
Willer
A
A
Fruehauf
S
S
Hochhaus
A
A
Hallek
M
M
Hehlmann
R
R
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002453:Cell Cycle / Q000502:physiology*; D002455:Cell Division; D002460:Cell Line; D019925:Cyclin A / Q000235:genetics; D005353:Fibronectins / Q000502:physiology*; D005434:Flow Cytometry; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000502:physiology*; D016193:G1 Phase; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology*; D016023:Integrins / Q000235:genetics; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016196:S Phase; D014162:Transfection
D019925:Cyclin A; D005353:Fibronectins; D016023:Integrins; D011993:Recombinant Fusion Proteins; D016044:Fusion Proteins, bcr-abl