S100A4 siRNA inhibits human pancreatic cancer cell invasion in vitro

Biomed Environ Sci. 2012 Aug;25(4):465-70. doi: 10.3967/0895-3988.2012.04.012.

Abstract

Objective: Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior.

Methods: Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase (MMP)-2, E-cadherin and thrombospondin (TSP)-1. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis.

Results: S100A4 mRNA expression was reduced to 17% after transfection with S100A4-siRNA, and protein expression had a similar trend. mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered.

Conclusion: S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing A100A4 can significantly contain the invasiveness of pancreatic cancer.

MeSH terms

  • Apoptosis / genetics
  • Apoptosis / physiology
  • Blotting, Western
  • Cadherins / genetics
  • Cadherins / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Humans
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism
  • Pancreatic Neoplasms / genetics
  • Pancreatic Neoplasms / metabolism*
  • RNA Interference
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • S100 Calcium-Binding Protein A4
  • S100 Proteins / genetics
  • S100 Proteins / metabolism*
  • Thrombospondin 1 / genetics
  • Thrombospondin 1 / metabolism

Substances

  • Cadherins
  • RNA, Small Interfering
  • S100 Calcium-Binding Protein A4
  • S100 Proteins
  • Thrombospondin 1
  • S100A4 protein, human
  • Matrix Metalloproteinase 2