Induction of tumorigenesis and metastasis by the murine orthologue of tumor protein D52

Jennifer D. Lewis, Laura A. Payton, Jill G. Whitford, Jennifer A. Byrne, David I Smith, LiBang Yang, Robert K. Bright

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Expression studies have consistently identified tumor protein D52 (TPD52) overexpression in tumor cells. Murine TPD52 (mD52) shares 86% identity with the human orthologue. To study a possible role for TPD52 in transformation, 3T3 fibroblasts were transfected with the full-length cDNA for mD52. Expression of mD52 was confirmed by reverse transcription-PCR (RT-PCR), real-time PCR, and Western blot analysis compared with 3T3 and vector-transfected 3T3 (3T3.V), and the resultant cell line was designated 3T3.mD52. At 4 weeks, 3T3.mD52 gained a 2-fold increase in growth rate, lost contact inhibition, and exhibited a marked phenotype change. Further characterization revealed an acquired ability for anchorage-independent cell growth. To determine whether 3T3.mD52 had become tumorigenic, naïve, healthy, immunocompetent syngeneic mice were inoculated subcutaneously with varying cell doses. Tumors measuring >1 cm2 were detected 60 days postinoculation with 3T3.mD52, and a 50% subcutaneous tumor incidence was obtained with as few as 5 × 105 3T3.mD52 cells. Remarkably, when lungs from 3T3.mD52 tumor-bearing mice were analyzed, numerous tumor nodules were observed, ranging from nodules less than 10 to nodules too numerous to count (inoculation with 1 × 105 and 5 × 106 cells, respectively). Further support for the metastatic capacity of 3T3.mD52 was the demonstration that transforming growth factor (TGF)-βR1 (receptor) expression decreased and TGF-β1 secretion increased in 3T3.mD52 compared with 3T3 controls. cDNA microarray analysis showed a gene expression pattern that further supported mD52-induced transformation and metastasis. Together, these data suggest that mD52 expression in 3T3 cells initiated cellular transformation, tumorigenesis, and progression to metastasis.

Original languageEnglish (US)
Pages (from-to)133-144
Number of pages12
JournalMolecular Cancer Research
Volume5
Issue number2
DOIs
StatePublished - Feb 2007

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Carcinogenesis
Neoplasm Metastasis
Neoplasms
Proteins
Transforming Growth Factors
Contact Inhibition
3T3 Cells
Growth Factor Receptors
Microarray Analysis
Growth
Oligonucleotide Array Sequence Analysis
Reverse Transcription
Real-Time Polymerase Chain Reaction
Complementary DNA
Fibroblasts
Western Blotting
Phenotype
Gene Expression
Cell Line
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

Cite this

Induction of tumorigenesis and metastasis by the murine orthologue of tumor protein D52. / Lewis, Jennifer D.; Payton, Laura A.; Whitford, Jill G.; Byrne, Jennifer A.; Smith, David I; Yang, LiBang; Bright, Robert K.

In: Molecular Cancer Research, Vol. 5, No. 2, 02.2007, p. 133-144.

Research output: Contribution to journalArticle

Lewis, Jennifer D. ; Payton, Laura A. ; Whitford, Jill G. ; Byrne, Jennifer A. ; Smith, David I ; Yang, LiBang ; Bright, Robert K. / Induction of tumorigenesis and metastasis by the murine orthologue of tumor protein D52. In: Molecular Cancer Research. 2007 ; Vol. 5, No. 2. pp. 133-144.
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abstract = "Expression studies have consistently identified tumor protein D52 (TPD52) overexpression in tumor cells. Murine TPD52 (mD52) shares 86{\%} identity with the human orthologue. To study a possible role for TPD52 in transformation, 3T3 fibroblasts were transfected with the full-length cDNA for mD52. Expression of mD52 was confirmed by reverse transcription-PCR (RT-PCR), real-time PCR, and Western blot analysis compared with 3T3 and vector-transfected 3T3 (3T3.V), and the resultant cell line was designated 3T3.mD52. At 4 weeks, 3T3.mD52 gained a 2-fold increase in growth rate, lost contact inhibition, and exhibited a marked phenotype change. Further characterization revealed an acquired ability for anchorage-independent cell growth. To determine whether 3T3.mD52 had become tumorigenic, na{\"i}ve, healthy, immunocompetent syngeneic mice were inoculated subcutaneously with varying cell doses. Tumors measuring >1 cm2 were detected 60 days postinoculation with 3T3.mD52, and a 50{\%} subcutaneous tumor incidence was obtained with as few as 5 × 105 3T3.mD52 cells. Remarkably, when lungs from 3T3.mD52 tumor-bearing mice were analyzed, numerous tumor nodules were observed, ranging from nodules less than 10 to nodules too numerous to count (inoculation with 1 × 105 and 5 × 106 cells, respectively). Further support for the metastatic capacity of 3T3.mD52 was the demonstration that transforming growth factor (TGF)-βR1 (receptor) expression decreased and TGF-β1 secretion increased in 3T3.mD52 compared with 3T3 controls. cDNA microarray analysis showed a gene expression pattern that further supported mD52-induced transformation and metastasis. Together, these data suggest that mD52 expression in 3T3 cells initiated cellular transformation, tumorigenesis, and progression to metastasis.",
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