title: Acquisition of interleukin-3 independence in FDC-P2 cells after transfection with the activated c-H-ras gene using a bovine papillomavirus-based plasmid vector.
abstract: Since the ras family of proto-oncogenes is supposed to be involved in leukemogenesis by point-mutational activation, we studied the effect of the activated ras gene on the growth of a murine interleukin-3 (IL-3)-dependent cell line, FDC-P2. The human activated c-H-ras gene was transfected into FDC-P2 cells by electroporation using a high-level expression vector, BMGhph, which contains a partial DNA sequence from bovine papillomavirus (BPV) and a hygromycin B (hmB)-resistant gene as a selectable marker. The transformed FDC-P2 cells showed a high incidence of IL-3-independent growth and tumorigenicity in nude mice. These clones did not express or secrete IL-3, suggesting the acquisition of IL-3 independence by a nonautocrine mechanism. The high incidence of autonomous growth may be due to the use of the BMG vector, because (1) the activated ras gene in pBR322 vector (pHs-49) was not so efficient in the induction of IL-3 independence, (2) the c-H-ras genome copies per cell increased in number up to about 50 copies by using the BMG vector, and (3) cotransfection with the activated ras gene and the BPV gene in separate plasmids partly enhanced the incidence of autonomous growth without increasing the copy number of the ras gene compared with transfection with the activated ras gene alone. The present study supports the idea that the activation of ras gene is an important step in malignant transformation of hematopoietic cells and suggests that the BPV gene products may cooperate with ras gene activation probably by affecting the cellular genes that may be involved in multistep leukemogenesis. The BMG vector may be useful to test the transforming ability of oncogenes whose oncogenic potential is relatively low.
authors.affiliation: Department of Hematology-Oncology, University of Tokyo, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Uemura
authors.forename: K
authors.initials: K
authors.lastname: Ozawa
authors.forename: A
authors.initials: A
authors.lastname: Tojo
authors.forename: K
authors.initials: K
authors.lastname: Takahashi
authors.forename: A
authors.initials: A
authors.lastname: Okano
authors.forename: H
authors.initials: H
authors.lastname: Karasuyama
authors.forename: K
authors.initials: K
authors.lastname: Tani
authors.forename: S
authors.initials: S
authors.lastname: Asano
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D052688:Bovine papillomavirus 1 / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D005091:Exons; D011905:Genes, ras; D005822:Genetic Vectors; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics / Q000494:pharmacology* / Q000502:physiology*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D007377:Interleukin-3; D012333:RNA, Messenger