The mouse int-2 gene exhibits basic fibroblast growth factor activity in a basic fibroblast growth factor-responsive cell line.

The int-2 protein is related to basic fibroblast growth factor (bFGF) by amino acid sequence homology. To assess its biological activity, we constructed retroviral vectors containing four variants of mouse int-2 complementary DNA under the transcriptional control of the beta-actin promoter and tested their effects on human SW13 adrenal cortical tumor cells. This cell line specifically requires bFGF, interleukin 1, or transforming growth factor e for anchorage-independent growth in soft agar. Despite encoding a signal sequence that should direct the protein to the secretory pathway, vectors containing unmodified int-2 complementary DNA, or a form optimized for translation initiation at the AUG codon, were incapable of inducing SW13 growth in soft agar. However, SW13 transfectants expressing a construct (pSP1), in which a mouse immunoglobulin signal peptide sequence is linked to the int-2 coding sequences, grew well in soft agar. The concentrated conditioned medium from these pSP1-transfected cells supported anchorage-independent growth of SW13 indicator cells and competed with bFGF for binding to receptors. Western blot analysis with an int-2-specific antiserum detected Mr 30,000-32,000 int-2 products in cell extracts and conditioned medium from pSP1-transfected clones, whereas the conditioned medium from these and other SW13 clones contained only low levels of bFGF as measured in a specific radioimmunoassay. These data suggest that the product of the int-2 gene can functionally replace bFGF in modulating the anchorage-independent growth of SW13 cells.

[1]  G. Peters,et al.  Subcellular fate of the lnt-2 oncoprotein is determined by choice of initiation codon , 1990, Nature.

[2]  R. Lupu,et al.  Autocrine growth stimulation by secreted Kaposi fibroblast growth factor but not by endogenous basic fibroblast growth factor. , 1990, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[3]  G. Peters,et al.  Detection and characterization of the fibroblast growth factor-related oncoprotein INT-2 , 1989, Molecular and cellular biology.

[4]  J. Halper,et al.  Modulation of growth of human carcinoma SW‐13 cells by heparin and growth factors , 1989, Journal of cellular physiology.

[5]  G. Merlo,et al.  Expression of int-2 mRNA in human tumors amplified at the int-2 locus. , 1989, Oncogene.

[6]  J. Rubin,et al.  Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. , 1989, Science.

[7]  D. Rifkin,et al.  Recent developments in the cell biology of basic fibroblast growth factor , 1989, The Journal of cell biology.

[8]  A. Sommer,et al.  Human basic fibroblast growth factor gene encodes four polypeptides: three initiate translation from non-AUG codons. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[9]  K. Takahashi,et al.  Growth stimulation of human breast epithelial cells by basic fibroblast growth factor in serum‐free medium , 1989, International journal of cancer.

[10]  J. Slack,et al.  Mesoderm-inducing properties of INT-2 and kFGF: two oncogene-encoded growth factors related to FGF. , 1989, Development.

[11]  G. Peters,et al.  Sequence organization of the human int-2 gene and its expression in teratocarcinoma cells. , 1989, Oncogene.

[12]  S. Nandi,et al.  Basic fibroblast growth factor stimulates the growth and inhibits casein accumulation in mouse mammary epithelial cells in vitro , 1989, Molecular and Cellular Endocrinology.

[13]  D. Birnbaum,et al.  Characterization of the HST-related FGF.6 gene, a new member of the fibroblast growth factor gene family. , 1989, Oncogene.

[14]  J. Lélias,et al.  High molecular mass forms of basic fibroblast growth factor are initiated by alternative CUG codons. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. Folkman,et al.  Basic fibroblast growth factor binds to subendothelial extracellular matrix and is released by heparitinase and heparin-like molecules. , 1989, Biochemistry.

[16]  M. Kozak.  The scanning model for translation: an update , 1989, The Journal of cell biology.

[17]  J. Fiddes,et al.  Addition of growth hormone secretion signal to basic fibroblast growth factor results in cell transformation and secretion of aberrant forms of the protein. , 1988, Oncogene.

[18]  M. Goldfarb,et al.  The human FGF-5 oncogene encodes a novel protein related to fibroblast growth factors , 1988, Molecular and cellular biology.

[19]  G. Martin,et al.  Four classes of mRNA are expressed from the mouse int‐2 gene, a member of the FGF gene family. , 1988, The EMBO journal.

[20]  G. Peters,et al.  Multiple RNAs expressed from the int‐2 gene in mouse embryonal carcinoma cell lines encode a protein with homology to fibroblast growth factors. , 1988, The EMBO journal.

[21]  D. Birnbaum,et al.  Chromosomal localization of the hst oncogene and its co-amplification with the int.2 oncogene in a human melanoma. , 1988, Oncogene.

[22]  A. McMahon,et al.  Expression of the FGF‐related proto‐oncogene int‐2 during gastrulation and neurulation in the mouse. , 1988, The EMBO journal.

[23]  G. Casey,et al.  Amplification of human int-2 in breast cancers and squamous carcinomas. , 1988, Oncogene.

[24]  G. Peters,et al.  Amplification of the int-2 gene in primary human breast tumors. , 1988, Oncogene research.

[25]  R. Weinberg,et al.  Basic fibroblast growth factor fused to a signal peptide transforms cells , 1988, Nature.

[26]  A. Hamburger,et al.  Stimulation of anchorage-independent growth of human tumor cells by interleukin 1. , 1987, Cancer research.

[27]  K. Miyagawa,et al.  Genomic sequence of hst, a transforming gene encoding a protein homologous to fibroblast growth factors and the int-2-encoded protein. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[28]  H. Moses,et al.  Purification and characterization of a novel transforming growth factor. , 1987, Cancer research.

[29]  M. Ittmann,et al.  An oncogene isolated by transfection of Kaposi's sarcoma DNA encodes a growth factor that is a member of the FGF family , 1987, Cell.

[30]  D. Gospodarowicz,et al.  Structural characterization and biological functions of fibroblast growth factor. , 1987, Endocrine reviews.

[31]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[32]  J. Folkman,et al.  Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[33]  H. Varmus,et al.  Two proto-oncogenes implicated in mammary carcinogenesis, int-1 and int-2, are independently regulated during mouse development. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[34]  S. O’Brien,et al.  Human endothelial cell growth factor: cloning, nucleotide sequence, and chromosome localization. , 1986, Science.

[35]  J. Fiddes,et al.  Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor. , 1986, Science.

[36]  D. Gospodarowicz,et al.  Molecular and biological characterization of fibroblast growth factor, an angiogenic factor which also controls the proliferation and differentiation of mesoderm and neuroectoderm derived cells. , 1986, Cell differentiation.

[37]  G. Peters,et al.  Sequence, topography and protein coding potential of mouse int‐2: a putative oncogene activated by mouse mammary tumour virus. , 1986, The EMBO journal.

[38]  G. Peters,et al.  Characterization and chromosome assignment of the human homolog of int-2, a potential proto-oncogene , 1986, Molecular and cellular biology.

[39]  D. Gospodarowicz,et al.  The identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells. , 1985, The Journal of biological chemistry.

[40]  G. Peters,et al.  Tumorigenesis by mouse mammary tumor virus: Proviral activation of a cellular gene in the common integration region int-2 , 1984, Cell.

[41]  H. Moses,et al.  Epithelial tissue-derived growth factor-like polypeptides. , 1983, Cancer research.

[42]  L. Hood,et al.  An immunoglobulin heavy chain variable region gene is generated from three segments of DNA: VH, D and JH , 1980, Cell.

[43]  Richard Axel,et al.  Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells , 1977, Cell.

[44]  A. Leibovitz,et al.  New human cancer cell culture lines. I. SW-13, small-cell carcinoma of the adrenal cortex. , 1973, Journal of the National Cancer Institute.

[45]  A. McMahon,et al.  Expression pattern of the FGF-related proto-oncogene int-2 suggests multiple roles in fetal development. , 1989, Development.

[46]  G. Merlo,et al.  The amplification unit on chromosome 11q13 in aggressive primary human breast tumors entails the bcl-1, int-2 and hst loci. , 1989, Oncogene.

[47]  T. Maciag,et al.  The heparin-binding (fibroblast) growth factor family of proteins. , 1989, Annual review of biochemistry.

[48]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[49]  G. Peters,et al.  Potential oncogene product related to growth factors , 1987, Nature.

[50]  G. Peters,et al.  On the Mechanism of Carcinogenesis by Mouse Mammary Tumor Virus , 1987 .