Clonal evolution in a myeloid cell line transformed to interleukin-3 independent growth by retroviral transduction and expression of p210bcr/abl.
Current evidence suggests that the expression of the tyrosine kinase p210bcr/abl in chronic myelogenous leukemia (CML) may directly induce the initial phase of granulocytic hyperplasia. However, the dysregulation of additional genes appears to be required for transition to the acute leukemic phase, as inferred by the appearance of recurrent secondary cytogenetic abnormalities in the majority of patients. To determine whether the expression of p210bcr/abl alone is responsible for this genetic instability, we introduced and expressed the bcr/abl gene from a retroviral vector in a clone of the interleukin-3 (IL-3) dependent myeloblastic 32D C13(G) cell line. Clonal and polyclonal cells transformed to IL-3 independent growth were observed for a period extending up to 6 months for changes in the expression of p210bcr/abl, cell proliferation, inhibition by prostaglandin E1 (PGE1), forskolin, and cyclic adenosine monophosphate (cAMP) analogues, regulation of the cell cycle, and karyotype. Whereas the properties of control vector infected 32D C13(G)' cells remained stable over time, cells expressing p210bcr/abl were phenotypically unstable. In cells expressing p210bcr/abl, we observed selective modulation of p210bcr/abl mRNA and protein expression, evolution from partial to full abrogation of IL-3 dependence, reduced serum requirements, increased cell proliferation, decreased inhibition by PGE1 and cAMP analogues, and the appearance of new structural and numerical chromosomal abnormalities with successive cell passages. These results indicate that expression of p210bcr/abl can directly predispose 32D C13(G)' cells to genetic instability, promotes the emergence of clones with an increased proliferative advantage, and may represent an in vitro model suitable for the study of mechanisms underlying progression to the acute leukemic phase in CML.
Laneuville
P
P
Department of Medicine, Royal Victoria Hospital, Montreal, Quebec, Canada.
Sun
G
G
Timm
M
M
Vekemans
M
M
D000782:Aneuploidy; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002453:Cell Cycle; D002455:Cell Division / Q000187:drug effects*; D002461:Cell Line, Transformed; D002999:Clone Cells; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D016044:Fusion Proteins, bcr-abl / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D007377:Interleukin-3 / Q000494:pharmacology*; D019868:Kanamycin Kinase; D007621:Karyotyping; D007700:Kinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D051379:Mice; D010770:Phosphotransferases / Q000235:genetics; D011401:Promoter Regions, Genetic; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012190:Retroviridae / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D004273:DNA, Neoplasm; D007377:Interleukin-3; D010770:Phosphotransferases; D013937:Thymidine Kinase; D019868:Kanamycin Kinase; D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl