Construction and characterization of recombinant adenoviruses expressing human BRCA1 or murine Brca1 genes

Mel Campbell, Olga N. Aprelikova, Riet Van Der Meer, Randall (Randy) Woltjer, Cindy J. Yee, Edison T. Liu, Roy A. Jensen

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Recombinant adenoviruses expressing human BRCA1 (AdBRCA1), murine Brca1 (AdBrca1), three clinically relevant human mutant BRCA1 proteins (t340, C61G, and 1853Stop), or a murine Brca1 C-terminal deletion mutant were constructed and evaluated in vitro. These recombinants were capable of transducing high-level transgene expression to a wide variety of cell lines in vitro. Three independent methods were utilized to monitor cell growth following transduction with these recombinants. High-level expression of either the human or mouse wild-type BRCA1 protein was incompatible with maximal levels of cell growth. AdBRCA1 transduction inhibited the outgrowth of several human breast and ovarian cell lines in colony formation assays. Flow cytometric analysis revealed an accumulation of the transduced cells in the G0/G1 phase of the cell cycle. This BRCA1-mediated accumulation of cells in G0/G1 was accompanied by an increase in the cellular level of hypophosphorylated pRB. Ad mutant BRCA1 t340, C61G, and 1853Stop viruses were impaired, to varying degrees, in their ability to transduce a growth-arrested state to the target cells. Using these same three criteria, overexpression of murine Brca1 by AdBrca1 was also capable of transducing a growth-arrested state to human cells. Deletion of the C-terminus of Brca1 diminished this activity. This panel of adenoviruses may be useful reagents as part of an approach to understand the function of BRCA1/Brca1 in normal breast and ovary and help to define the tumor suppressor defect(s) conferred by clinical BRCA1 mutations in breast and ovarian cell tumorigenesis.

Original languageEnglish (US)
Pages (from-to)231-239
Number of pages9
JournalCancer Gene Therapy
Volume8
Issue number3
StatePublished - 2001
Externally publishedYes

Fingerprint

Human Adenoviruses
Genes
Breast
Growth
BRCA1 Protein
Cell Line
Cell Cycle Resting Phase
G1 Phase
Mutant Proteins
Transgenes
Adenoviridae
Ovary
Cell Cycle
Carcinogenesis
Viruses
Mutation

Keywords

  • Adenoviruses
  • Cell cycle
  • Growth inhibition
  • Tumor suppressor

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Campbell, M., Aprelikova, O. N., Van Der Meer, R., Woltjer, R. R., Yee, C. J., Liu, E. T., & Jensen, R. A. (2001). Construction and characterization of recombinant adenoviruses expressing human BRCA1 or murine Brca1 genes. Cancer Gene Therapy, 8(3), 231-239.

Construction and characterization of recombinant adenoviruses expressing human BRCA1 or murine Brca1 genes. / Campbell, Mel; Aprelikova, Olga N.; Van Der Meer, Riet; Woltjer, Randall (Randy); Yee, Cindy J.; Liu, Edison T.; Jensen, Roy A.

In: Cancer Gene Therapy, Vol. 8, No. 3, 2001, p. 231-239.

Research output: Contribution to journalArticle

Campbell, M, Aprelikova, ON, Van Der Meer, R, Woltjer, RR, Yee, CJ, Liu, ET & Jensen, RA 2001, 'Construction and characterization of recombinant adenoviruses expressing human BRCA1 or murine Brca1 genes', Cancer Gene Therapy, vol. 8, no. 3, pp. 231-239.
Campbell, Mel ; Aprelikova, Olga N. ; Van Der Meer, Riet ; Woltjer, Randall (Randy) ; Yee, Cindy J. ; Liu, Edison T. ; Jensen, Roy A. / Construction and characterization of recombinant adenoviruses expressing human BRCA1 or murine Brca1 genes. In: Cancer Gene Therapy. 2001 ; Vol. 8, No. 3. pp. 231-239.
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