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Inhibiting effect of murine double minute-2 oncogene on apoptosis induced by retroviral-mediated wild-type p53.

Publication ,  Journal Article
Guo, H; Liu, T; Gao, J
Published in: Chin Med J (Engl)
December 1998

OBJECTIVE: To investigate the effects of wild-type p53 (wtp53) on inducing apoptosis by restoring wtp53 expression in pancreatic adenocarcinoma cell line (PC-2) which contains mutant p53, and the interaction between murine double minute-2 (MDM2) and wtp53 in pancreatic adenocarcinoma. METHODS: A recombinant retroviral vector expressing wild-type p53 was constructed and packaged by packaging cell line PA317 cells using calcium phosphate coprecipitation method. The supernatant of the packaged cells PA317 was used to transfect the pancreatic carcinoma cell line (PC-2), then a transformed cell line PC-2/swtp53 was established. The recombinant vector pCMV-MDM2 was transduced into PC-2/swtp53 cell line by lipofectin-mediated method, a double transfected cell line (PC-2/swtp53/pCMV-MDM2) was formed. To determine the integration and expression of exogenous wtp53 gene in the transfected cells, polymerase chain reaction (PCR), Western blot and immunoprecipitation analyses were performed. Apoptosis was analyzed by means of flow cytometry, in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) analysis and DNA agarose gel electrophoresis. RESULTS: Transduction with the retroviral vector resulted in integration and expression of wtp53 gene in host cells. The apoptotic cells in PC-2/swtp53 and PC-2/swtp53/pCMV-neo cell lines were 12.1%-12.9%, while the double transfected cell line, PC-2/swtp53/pCMV-MDM2, showed less (3.2%) apoptotic cells than its parent cell lines. CONCLUSIONS: Restoration of expression of wild-type p53 with a retroviral vector can increase programmed cell death of pancreatic adenocarcinoma cells (PC-2) containing mutant p53. The overexpression of MDM2 protein has a negative regulating role in wtp53-induced apoptosis in PC-2 cell.

Duke Scholars

Published In

Chin Med J (Engl)

ISSN

0366-6999

Publication Date

December 1998

Volume

111

Issue

12

Start / End Page

1102 / 1106

Location

China

Related Subject Headings

  • Tumor Cells, Cultured
  • Transfection
  • Retroviridae
  • Proto-Oncogene Proteins c-mdm2
  • Proto-Oncogene Proteins
  • Pancreatic Neoplasms
  • Oncology & Carcinogenesis
  • Oncogenes
  • Nuclear Proteins
  • Neoplasm Proteins
 

Citation

APA
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ICMJE
MLA
NLM
Guo, H., Liu, T., & Gao, J. (1998). Inhibiting effect of murine double minute-2 oncogene on apoptosis induced by retroviral-mediated wild-type p53. Chin Med J (Engl), 111(12), 1102–1106.
Guo, H., T. Liu, and J. Gao. “Inhibiting effect of murine double minute-2 oncogene on apoptosis induced by retroviral-mediated wild-type p53.Chin Med J (Engl) 111, no. 12 (December 1998): 1102–6.
Guo, H., et al. “Inhibiting effect of murine double minute-2 oncogene on apoptosis induced by retroviral-mediated wild-type p53.Chin Med J (Engl), vol. 111, no. 12, Dec. 1998, pp. 1102–06.

Published In

Chin Med J (Engl)

ISSN

0366-6999

Publication Date

December 1998

Volume

111

Issue

12

Start / End Page

1102 / 1106

Location

China

Related Subject Headings

  • Tumor Cells, Cultured
  • Transfection
  • Retroviridae
  • Proto-Oncogene Proteins c-mdm2
  • Proto-Oncogene Proteins
  • Pancreatic Neoplasms
  • Oncology & Carcinogenesis
  • Oncogenes
  • Nuclear Proteins
  • Neoplasm Proteins