Dominant negative Met reduces tumorigenicity-metastasis and increases tubule formation in mammary cells

Activation of the Met tyrosine kinase growth factor receptor by its ligand HGF/SF has been shown to increase in vitro invasiveness in epithelial cell lines. To study the effect of Met-HGF/SF signaling in breast cancer cells, we transfected met, hgf/sf and dominant negative (DN) forms of met into the poorly differentiated metastatic murine mammary adenocarcinoma cell line DA3. These cells express moderate levels of endogenous Met, which is rapidly phosphorylated in response to HGF/SF treatment. Met+hgf/sf transfection results in significantly increased tumorigenic and metastatic activity in vivo accompanied by reduced tubule formation. DA3 cells transfected with DN forms of Met (DN-DA3) exhibit reduced Met phosphorylation following exposure to HGF/SF. Furthermore, as compared to the parental cells, the DN-DA3 cells exhibit diminished in vitro scattering and invasiveness, while in vivo they display greatly reduced tumorigenicity and spontaneous metastasis. Tumors emanating from DN-DA3 cells injected to BALB/C mice are highly differentiated and display extensive tubule formation. These results suggest that Met-HGF/SF signaling is a determining factor in the delicate balance between differentiation/tubule formation and tumorigenicity-metastasis.

[1]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[2]  E T BELL,et al.  The Diseases of the Breast , 1925, Nature.

[3]  C. D. HAAGENSEN,et al.  Diseases of the Breast , 1972 .

[4]  I. Pastan,et al.  The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Michael Stoker,et al.  Scatter factor is a fibroblast-derived modulator of epithelial cell mobility , 1987, Nature.

[6]  Y. Yarden,et al.  Epidermal growth factor induces rapid, reversible aggregation of the purified epidermal growth factor receptor. , 1987, Biochemistry.

[7]  G. V. Vande Woude,et al.  Characterization of the TPR-MET oncogene p65 and the MET protooncogene p140 protein-tyrosine kinases. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[8]  K. Tashiro,et al.  Molecular cloning and expression of human hepatocyte growth factor , 1989, Nature.

[9]  Joseph Schlessinger,et al.  Signal transduction by receptors with tyrosine kinase activity , 1990, Cell.

[10]  M. Kirschner,et al.  Expression of a dominant negative mutant of the FGF receptor disrupts mesoderm formation in xenopus embryos , 1991, Cell.

[11]  I. Bièche,et al.  Loss of heterozygosity on chromosome 7q and aggressive primary breast cancer , 1992, The Lancet.

[12]  L. Naldini,et al.  Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motility and growth , 1992, The Journal of cell biology.

[13]  G. V. Vande Woude,et al.  The met proto-oncogene receptor and lumen formation. , 1992, Science.

[14]  T. Nakamura,et al.  Tumorigenicity of the met proto-oncogene and the gene for hepatocyte growth factor , 1992, Molecular and cellular biology.

[15]  G. Rodrigues,et al.  Dimerization mediated through a leucine zipper activates the oncogenic potential of the met receptor tyrosine kinase , 1993, Molecular and cellular biology.

[16]  P. Comoglio,et al.  Expression of the c-Met/HGF receptor in human melanocytic neoplasms: demonstration of the relationship to malignant melanoma tumour progression. , 1993, British Journal of Cancer.

[17]  E Medico,et al.  Transfer of motogenic and invasive response to scatter factor/hepatocyte growth factor by transfection of human MET protooncogene. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  G. V. Vande Woude,et al.  The Met proto-oncogene mesenchymal to epithelial cell conversion. , 1994, Science.

[19]  A. Bardelli,et al.  A multifunctional docking site mediates signaling and transformation by the hepatocyte growth factor/scatter factor receptor family , 1994, Cell.

[20]  M. Bhargava,et al.  Scatter factor modulates the metastatic phenotype of the EMT6 mouse mammary tumor , 1994, International journal of cancer.

[21]  G. Cunha.  Role of mesenchymal‐epithelial interactions in normal and abnormal development of the mammary gland and prostate , 1994, Cancer.

[22]  P Zola,et al.  Overexpression of the MET/HGF receptor in ovarian cancer , 1994, International journal of cancer.

[23]  G. V. Vande Woude,et al.  Invasiveness and metastasis of NIH 3T3 cells induced by Met-hepatocyte growth factor/scatter factor autocrine stimulation. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Carmen Birchmeier,et al.  Essential role for the c-met receptor in the migration of myogenic precursor cells into the limb bud , 1995, Nature.

[25]  Remo Guidieri.  Res , 1995, RES: Anthropology and Aesthetics.

[26]  M. Sharpe,et al.  Scatter factor/hepatocyte growth factor is essential for liver development , 1995, Nature.

[27]  S. Ethier.  Growth factor synthesis and human breast cancer progression. , 1995, Journal of the National Cancer Institute.

[28]  C. Benz,et al.  Gene diagnostics provide new insights into breast cancer prognosis and therapy. , 1995, Gene.

[29]  G. V. Vande Woude,et al.  The Met-HGF/SF autocrine signaling mechanism is involved in sarcomagenesis. , 1995, EXS.

[30]  W. Birchmeier,et al.  Mutation of juxtamembrane tyrosine residue 1001 suppresses loss-of-function mutations of the met receptor in epithelial cells. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[31]  L. Orci,et al.  Modulation of hepatocyte growth factor and c-met in the rat mammary gland during pregnancy, lactation, and involution. , 1995, Experimental cell research.

[32]  A. Purushotham,et al.  Expression and loss of heterozygosity of c‐met proto‐oncogene in primary breast cancer , 1995, Journal of surgical oncology.

[33]  Tetsuo Noda,et al.  Placental defect and embryonic lethality in mice lacking hepatocyte growth factor/scatter factor , 1995, Nature.

[34]  W. Birchmeier,et al.  Sequential requirement of hepatocyte growth factor and neuregulin in the morphogenesis and differentiation of the mammary gland , 1995, The Journal of cell biology.

[35]  R. Cardiff,et al.  Transgenic expression of tpr-met oncogene leads to development of mammary hyperplasia and tumors. , 1996, The Journal of clinical investigation.

[36]  G. V. Vande Woude,et al.  Autocrine hepatocyte growth factor/scatter factor-Met signaling induces transformation and the invasive/metastastic phenotype in C127 cells. , 1996, Oncogene.

[37]  G. V. Vande Woude,et al.  Enhanced tumorigenicity and invasion-metastasis by hepatocyte growth factor/scatter factor-met signalling in human cells concomitant with induction of the urokinase proteolysis network , 1996, Molecular and cellular biology.

[38]  Germline and somatic mutations in the tyrosine kinase domain of the MET proto-oncogene in papillary renal carcinomas , 1997, Nature Genetics.

[39]  R. Sharp,et al.  Diverse tumorigenesis associated with aberrant development in mice overexpressing hepatocyte growth factor/scatter factor. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[40]  R. Kramer,et al.  Expression of the C‐Met/HGF receptor in human breast carcinoma: Correlation with tumor progression , 1997, International journal of cancer.

[41]  S. Schnitt,et al.  Expression of scatter factor and c‐met receptor in benign and malignant breast tissue , 1997 .

[42]  L. Schmidt,et al.  Activating mutations for the met tyrosine kinase receptor in human cancer. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[43]  E. Rimm,et al.  Expression of c‐met is a strong independent prognostic factor in breast carcinoma , 1998, Cancer.

[44]  A. Bardelli,et al.  Induction of epithelial tubules by growth factor HGF depends on the STAT pathway , 1998, Nature.

[45]  N. Hacohen,et al.  sprouty Encodes a Novel Antagonist of FGF Signaling that Patterns Apical Branching of the Drosophila Airways , 1998, Cell.