Autocrine transformation of human hematopoietic cells after transfection with an activated granulocyte/macrophage colony stimulating factor gene

Cytokines Cell Mol Ther. 1997 Sep;3(3):159-68.

Abstract

The effects of constitutive cytokine gene expression on the growth-factor-dependence of the human erythroleukemic TF-1 cell line have been determined. TF-1 cells normally require the presence of exogenous cytokines to proliferate in vitro. TF-1 cells were transfected with constructs containing either the germline granulocyte-macrophage colony-stimulating factor (GM-CSF) gene or the GM-CSF gene linked to the Moloney murine leukemia virus (Mo-MuLV) long terminal repeat. The Mo-MuLV-LTR, which contains a strong transcriptional enhancer, was added to stimulate the constitutive expression of the GM-CSF gene. Transfection with the germline GM-CSF gene did not abrogate the cytokine dependence of TF-1 cells, indicating that inheritance of an extra copy did not result in sufficient GM-CSF expression to abrogate cytokine dependence. In contrast, transfection with the LTR-modified GM-CSF gene resulted in the isolation of cells that proliferated in the absence of exogenous GM-CSF. The LTR increased nascent transcription and accumulation of GM-CSF mRNA transcripts, which had a normal half-life. This increase in GM-CSF expression led to secretion of sufficient GM-CSF to support the growth of the parental TF-1 cells. These results indicate that the deregulated expression of human cytokine genes induced by certain retroviral LTRs can result in their conversion into hematopoietic-specific oncogenes. These modified human cell lines provide a model to investigate autocrine transformation and therapy of acute myelogenous leukemia as well as other hematopoietic disorders.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Autocrine Communication*
  • Blotting, Northern
  • Blotting, Southern
  • Cell Division / drug effects
  • Cell Transformation, Neoplastic*
  • Cytokines / genetics
  • Cytokines / pharmacology
  • Enhancer Elements, Genetic
  • Gene Expression Regulation, Neoplastic
  • Genes, Viral / genetics
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics*
  • Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Growth Substances / pharmacology
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Leukemia, Erythroblastic, Acute
  • Moloney murine leukemia virus / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Repetitive Sequences, Nucleic Acid / genetics
  • Transcription, Genetic / genetics
  • Transfection*
  • Tumor Cells, Cultured

Substances

  • Cytokines
  • Growth Substances
  • RNA, Messenger
  • Granulocyte-Macrophage Colony-Stimulating Factor