Inhibition of TGF-beta receptor I by siRNA suppresses the motility and invasiveness of T24 bladder cancer cells via modulation of integrins and matrix metalloproteinase

Int Urol Nephrol. 2010 Jun;42(2):315-23. doi: 10.1007/s11255-009-9620-3. Epub 2009 Aug 8.

Abstract

Background: Urinary bladder transitional-cell carcinoma is still challenging because the mechanisms underlying the tumor progression are still largely unknown. Transforming growth factor beta1 (TGF-beta1) is considered a crucial molecule in the tumorigenesis of urinary bladder carcinoma. Many studies have indicated that it is also associated with epithelial-mesenchymal transition, angiogenesis, migration and metastases in many types of malignant tumors.

Materials and methods: We blocked the TGF-beta signal pathway in T24 human bladder cancer cells with a siRNA (TsiRNA), which targets the TGF-beta type I receptor and evaluated the effects of TGF-beta1 and TsiRNA on the cell motility and invasiveness by Matrigel migration assay, wound-healing assay and Matrigel invasion assay. RT-PCR and Western blotting analysis were used to examine the effects of TGF-beta1 and TsiRNA on the expression of TGFBRI and genes, which are related to tumor migration and invasion.

Results: While exogenous TGF-beta1 enhanced the migration and invasion of T24 cells, TsiRNA significantly suppressed them. RT-PCR and Western blotting analysis revealed that TsiRNA could downregulate both the expression of alpha3, beta1 and alpha2 integrin subunits and the activity of matrix metalloproteinase 9 enhanced by exogenous TGF-beta1.

Conclusion: Our study suggested that inhibition of TGF-beta1 signaling pathway by siRNA could be beneficial in the treatment of patients with metastatic bladder cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Transitional Cell / genetics
  • Carcinoma, Transitional Cell / pathology*
  • Cell Movement
  • Down-Regulation
  • Humans
  • Integrins / physiology*
  • Matrix Metalloproteinases / physiology*
  • Neoplasm Invasiveness
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / genetics*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology*
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptors, Transforming Growth Factor beta / antagonists & inhibitors*
  • Receptors, Transforming Growth Factor beta / genetics*
  • Signal Transduction
  • Tumor Cells, Cultured
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / pathology*

Substances

  • Integrins
  • RNA, Small Interfering
  • Receptors, Transforming Growth Factor beta
  • Protein Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type I
  • Matrix Metalloproteinases