title: BCR/ABL induces multiple abnormalities of cytoskeletal function.
abstract: The BCR/ABL oncogene causes human chronic myelogenous leukemia (CML), a myeloproliferative disease characterized by massive expansion of hematopoietic progenitor cells and cells of the granulocyte lineage. When transfected into murine hematopoietic cell lines, BCR/ABL causes cytokine-independence and enhances viability. There is also growing evidence that p210(BCR/ABL) affects cytoskeletal structure. p210(BCR/ABL) binds to actin, and several cytoskeletal proteins are tyrosine phosphorylated by this oncoprotein. Also, at least one aspect of cytoskeletal function is abnormal, in that the affinity of beta1 integrins for fibronectin is altered in CML cells. However, isolated changes in beta1 integrin function would be unlikely to explain the clinical phenotype of CML. We used time-lapse video microscopy to study cell motility and cell morphology on extracellular cell matrix protein-coated surfaces of a series of cell lines before and after transformation by BCR/ABL. BCR/ABL was associated with a striking increase in spontaneous motility, membrane ruffling, formation of long actin extensions (filopodia) and accelerated the rate of protrusion and retraction of pseudopodia on fibronectin-coated surfaces. Also, while untransformed cells were sessile for long periods, BCR/ABL-transformed cells exhibited persistent motility, except for brief periods during cell division. Using cell lines transformed by a temperature-sensitive mutant of BCR/ABL, these kinetic abnormalities of cytoskeletal function were shown to require BCR/ABL tyrosine kinase activity. Similar abnormalities of cytoskeletal function on fibronectin-coated surfaces were observed when hematopoietic progenitor cells purified by CD34 selection from patients with CML were compared with CD34 positive cells from normal individuals. Interestingly, alpha-interferon treatment was found to slowly revert the abnormal motility phenotype of BCR/ABL-transformed cells towards normal. The increase in spontaneous motility and other defects of cytoskeletal function described here will be useful biological markers of the functional effects of BCR/ABL in hematopoietic cells.
authors.affiliation: Division of Hematologic Malignancies, Dana-Farber Cancer Institute and Harvard Medical School, Boston, Massachusetts 02115, USA. Ravi_Salgia@dfci.harvard.edu
authors.forename: R
authors.initials: R
authors.lastname: Salgia
authors.forename: J L
authors.initials: JL
authors.lastname: Li
authors.forename: D S
authors.initials: DS
authors.lastname: Ewaniuk
authors.forename: W
authors.initials: W
authors.lastname: Pear
authors.forename: E
authors.initials: E
authors.lastname: Pisick
authors.forename: S A
authors.initials: SA
authors.lastname: Burky
authors.forename: T
authors.initials: T
authors.lastname: Ernst
authors.forename: M
authors.initials: M
authors.lastname: Sattler
authors.forename: L B
authors.initials: LB
authors.lastname: Chen
authors.forename: J D
authors.initials: JD
authors.lastname: Griffin
mesh_terms: D016475:3T3 Cells; D000199:Actins / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D001853:Bone Marrow / Q000473:pathology; D002448:Cell Adhesion; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002471:Cell Transformation, Neoplastic; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000473:pathology*; D005109:Extracellular Matrix / Q000502:physiology; D016044:Fusion Proteins, bcr-abl / Q000096:biosynthesis / Q000235:genetics*; D016313:Genes, abl; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000473:pathology; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D007700:Kinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000473:pathology; D051379:Mice; D018715:Microscopy, Video; D009857:Oncogenes; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012016:Reference Values; D013696:Temperature; D014162:Transfection
chemical_list: D000199:Actins; D015703:Antigens, CD; D018952:Antigens, CD34; D003598:Cytoskeletal Proteins; D016898:Interferon-alpha; D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl