Skip to main content
Journal cover image

Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells.

Publication ,  Journal Article
Mithani, SK; Balch, GC; Shiou, S-R; Whitehead, RH; Datta, PK; Beauchamp, RD
Published in: J Surg Res
April 2004

BACKGROUND: Smad proteins play a key role in TGF-beta signaling that regulates cell proliferation, differentiation, and apoptosis. Mice deficient in Smad3 develop colonic adenocarcinoma. MATERIALS AND METHODS: We developed a Smad3-deficient colonocyte cell line that was used to study TGF-beta-mediated growth inhibition and induction of apoptosis was compared to young adult mouse colonocyte (YAMC) control cells. Growth inhibition was assessed by cell count and ((3)H)-thymidine incorporation assay. Transcriptional response to TGF-beta was measured by transfecting the reporters p3TP-Lux and p(CAGA)(9)-MLP-luc. TGF-beta-induced apoptosis was assessed using ELISA and Hoechst staining. Mediators of cell-cycle arrest and apoptosis were assayed by Western blot. RESULTS: Smad3-/- cells were resistant to TGF-beta-mediated growth inhibition compared to control cells. Ninety-eight percent of cell count growth inhibition observed in YAMC cells, while 34% inhibition was observed in Smad3-/- cells after TGF-beta treatment. ((3)H)-thymidine incorporation was inhibited by 61% in YAMC cells, while Smad3-/- cells showed 25% inhibition after TGF-beta treatment. Smad3-/- cells were deficient in luciferase reporter induction by TGF-beta. TGF-beta induced apoptosis 8-fold in YAMC cells, but had no effect on apoptosis in Smad3-/- cells. p21(Cip11) and PAI-1 are induced in YAMC cells by TGF-beta, but unchanged in Smad3-/- cells. TGF-beta decreases cyclin D1 levels in YAMC cells but does not affect levels in Smad3-/- cells. CONCLUSIONS: Our findings suggest that the loss of Smad3 contributes to resistance of TGF-beta growth inhibition and apoptosis in colonic epithelium. This may represent a mechanism by which cells are able to escape antiproliferative controls and embark on a pathway toward neoplasia.

Duke Scholars

Published In

J Surg Res

DOI

ISSN

0022-4804

Publication Date

April 2004

Volume

117

Issue

2

Start / End Page

296 / 305

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Trans-Activators
  • Surgery
  • Smad3 Protein
  • Promoter Regions, Genetic
  • Mutation
  • Mice, Transgenic
  • Mice
  • Intestinal Mucosa
  • Growth Inhibitors
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Mithani, S. K., Balch, G. C., Shiou, S.-R., Whitehead, R. H., Datta, P. K., & Beauchamp, R. D. (2004). Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells. J Surg Res, 117(2), 296–305. https://doi.org/10.1016/S0022-4804(03)00335-4
Mithani, Suhail K., Glen C. Balch, Sheng-Ru Shiou, Robert H. Whitehead, Pran K. Datta, and R Daniel Beauchamp. “Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells.J Surg Res 117, no. 2 (April 2004): 296–305. https://doi.org/10.1016/S0022-4804(03)00335-4.
Mithani SK, Balch GC, Shiou S-R, Whitehead RH, Datta PK, Beauchamp RD. Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells. J Surg Res. 2004 Apr;117(2):296–305.
Mithani, Suhail K., et al. “Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells.J Surg Res, vol. 117, no. 2, Apr. 2004, pp. 296–305. Pubmed, doi:10.1016/S0022-4804(03)00335-4.
Mithani SK, Balch GC, Shiou S-R, Whitehead RH, Datta PK, Beauchamp RD. Smad3 has a critical role in TGF-beta-mediated growth inhibition and apoptosis in colonic epithelial cells. J Surg Res. 2004 Apr;117(2):296–305.
Journal cover image

Published In

J Surg Res

DOI

ISSN

0022-4804

Publication Date

April 2004

Volume

117

Issue

2

Start / End Page

296 / 305

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Trans-Activators
  • Surgery
  • Smad3 Protein
  • Promoter Regions, Genetic
  • Mutation
  • Mice, Transgenic
  • Mice
  • Intestinal Mucosa
  • Growth Inhibitors