Effects of gastrin 17 on beta-catenin/Tcf-4 pathway in Colo320WT colon cancer cells

World J Gastroenterol. 2006 Dec 14;12(46):7482-7. doi: 10.3748/wjg.v12.i46.7482.


Aim: To explore the effect of gastrin 17 (G17) on beta-catenin/T cell factor-4 (Tcf-4) signaling in colonic cancer cell line Colo320WT.

Methods: The pCR3.1/GR plasmid, which expresses gastrin receptor, cholecystokinin-2 receptor (CCK-2R), was transfected into a colonic cancer cell line Colo320 by Lipofectamine (TM)2000 and the stably expressing CCK-2R clones were screened by G418. The expression levels of gastrin receptor in the Colo320 and the transfected Colo320WT cell line were assayed by RT-PCR. Colo320WT cells were treated with G17 in a time-dependent manner (0, 1, 6, 12, 24 and 48 h), then with L365,260 (Gastrin(17) receptor blocker) for 30 min, and with G17 again for 12 h or L365,260 for 12 h. Expression levels of beta-catenin in a TX-100 soluble fraction and TX-100 insoluble fraction of Colo320WT cells treated with G17 were detected by co-immuniprecipation and Western blot. Immunocytochemistry was used to examine the distribution of beta-catenin in CoLoWT320 cells. Expression levels of c-myc and cyclin D1 in Colo320WT cells treated with G17 were assayed by Western blot.

Results: Expression levels of beta-catenin in the TX-100 solution fraction decreased apparently in a time-dependent fashion and reached the highest level after G17 treatment for 12 h, while expression levels of beta-catenin in the TX-100 insoluble fraction were just on the contrary. Immunocytochemistry showed that beta-catenin was translocated from the cell membranes into the cytoplasm and nucleus under G17 treatment. Expression levels of c-myc and cyclin D1 in the G17-treated Colo320WT cells were markedly higher compared to the untreated Colo320WT cells. In addition, the aforementioned G17-stimulated responses were blocked by L365,260.

Conclusion: Gastrin17 activates beta-catenin/Tcf-4 signaling in Colo320WT cells, thereby leading to over-expression of c-myc and cyclin D1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line, Tumor
  • Colonic Neoplasms / drug therapy*
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism*
  • Cyclin D
  • Cyclins / metabolism
  • DNA, Complementary / genetics
  • Gastrins / pharmacology*
  • Humans
  • Proto-Oncogene Proteins c-myc / metabolism
  • Receptor, Cholecystokinin B / genetics
  • Receptor, Cholecystokinin B / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • TCF Transcription Factors / metabolism*
  • Transcription Factor 7-Like 2 Protein
  • Transfection
  • beta Catenin / metabolism*


  • CTNNB1 protein, human
  • Cyclin D
  • Cyclins
  • DNA, Complementary
  • Gastrins
  • MYC protein, human
  • Proto-Oncogene Proteins c-myc
  • Receptor, Cholecystokinin B
  • Recombinant Proteins
  • TCF Transcription Factors
  • TCF7L2 protein, human
  • Transcription Factor 7-Like 2 Protein
  • beta Catenin
  • gastrin 17