Molecular consequences of silencing mutant K-ras in pancreatic cancer cells

Justification for K-ras-directed therapy

Jason B. Fleming, Guo Liang Shen, Shane E. Holloway, Mishel Davis, Rolf A. Brekken

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

Mutation of the K-ras gene is an early event in the development of pancreatic adenocarcinoma and, therefore, RNA interference (RNAi) directed toward mutant K-ras could represent a novel therapy. In this study, we examine the phenotypic and molecular consequences of exposure of pancreatic tumor cells to mutant-specific K-ras small interfering RNA. Specific reduction of activated K-ras via RNAi in Panc-1 and MiaPaca-2 cells resulted in cellular changes consistent with a reduced capacity to form malignant tumors. These changes occur through distinct mechanisms but likely reflect an addiction of each cell line to oncogene stimulation. Both cell lines show reduced proliferation after K-ras RNAi, but only MiaPaca-2 cells showed increased apoptosis. Both cell lines showed reduced migration after K-ras knockdown, but changes in integrin levels were not consistent between the cell lines. Both cell lines showed alteration of the level of GLUT-1, a metabolism-associated gene that is downstream of c-myc, with Panc-1 cells demonstrating decreased GLUT-1 levels, whereas MiaPaca-2 cells showed increased levels of expression after K-ras knockdown. Furthermore, after K-ras RNAi, there was a reduction in angiogenic potential of both Panc-1 and MiaPaca-2 cells. Panc-1 cells increased the level of expression of thrombospondin-1, an endogenous inhibitor of angiogenesis, whereas MiaPaca-2 cells decreased the production of vascular endothelial growth factor, a primary stimulant of angiogenesis in pancreatic tumors. We have found that silencing mutant K-ras through RNAi results in alteration of tumor cell behavior in vitro and suggests that targeting mutant K-ras specifically might be effective against pancreatic cancer in vivo.

Original languageEnglish (US)
Pages (from-to)413-423
Number of pages11
JournalMolecular Cancer Research
Volume3
Issue number7
DOIs
StatePublished - Jul 2005

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Pancreatic Neoplasms
RNA Interference
Cell Line
Therapeutics
Neoplasms
Thrombospondin 1
Angiogenesis Inhibitors
ras Genes
Oncogenes
Integrins
Vascular Endothelial Growth Factor A
Small Interfering RNA
Adenocarcinoma
Apoptosis
Mutation
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

Cite this

Molecular consequences of silencing mutant K-ras in pancreatic cancer cells : Justification for K-ras-directed therapy. / Fleming, Jason B.; Shen, Guo Liang; Holloway, Shane E.; Davis, Mishel; Brekken, Rolf A.

In: Molecular Cancer Research, Vol. 3, No. 7, 07.2005, p. 413-423.

Research output: Contribution to journalArticle

Fleming, Jason B. ; Shen, Guo Liang ; Holloway, Shane E. ; Davis, Mishel ; Brekken, Rolf A. / Molecular consequences of silencing mutant K-ras in pancreatic cancer cells : Justification for K-ras-directed therapy. In: Molecular Cancer Research. 2005 ; Vol. 3, No. 7. pp. 413-423.
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