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Molecular Consequences of Silencing Mutant K-ras in Pancreatic Cancer Cells: Justification for K-ras–Directed Therapy
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.
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“Silencing mutant KRAS in human pancreatic cancer cells using specific siRNA effectively reduces KRAS expression, leading to a dose-dependent decrease in KRAS protein levels.”
From Significance of TP53, CDKN2A, SMAD4 and KRAS in Pancreatic Cancer (2024) · cites this paper as [125]
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“In KRAS mutant pancreatic cancer cells, acute silencing of KRAS via RNAi or CRISPR results in cell cycle arrest, apoptosis and suppression of anchorage-independent proliferation in vitro, and attenuated tumor growth and metastasis in xenograft models (66–68).”
From KRAS mutation in pancreatic cancer (2021) · cites this paper as [66]
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Metadata
- Authors
- Fleming, J. B., Shen, G.-L., Holloway, S. E., Davis, M., & Brekken, R. A.
- Affiliations
- 1Division of Surgical Oncology, Department of Surgery, and Hamon Center for Therapeutic Oncology Research and
- The University of Texas Southwestern Medical Center (US)
- Venue
- Molecular Cancer Research
- Published
- 2005-07-01
- Volume
- 3
- Issue
- 7
- Pages
- 413-423
- ISSN
- 1541-7786 (ISSN-L)
- 1557-3125
- OpenAlex
- W2154164444
- Cited by
- 176
- References
- 63
- Citations by year
- 2026: 3 · 2024: 2 · 2023: 4 · 2022: 7 · 2021: 10 · 2020: 8 · 2019: 4 · 2018: 3 · 2017: 11 · 2016: 5 · 2015: 9 · 2014: 18 · 2013: 11 · 2012: 11
- FWCI
- 5.75
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Molecular Cancer Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Cancer Institute (R21 CA10669)
- Topics
- Health Sciences › Medicine › Oncology › Pancreatic and Hepatic Oncology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
- Keywords
- mutant KRAS, pancreatic ductal adenocarcinoma, RNA interference, siRNA, oncogene addiction, cell proliferation, apoptosis, cell migration, integrins, GLUT1, c-Myc, angiogenesis, thrombospondin-1, vascular endothelial growth factor, PANC-1 cells, MIA PaCa-2 cells
- MeSH terms
- Adenocarcinoma, Animals, Cell Movement, Humans, Immunohistochemistry, Mutation, Pancreatic Neoplasms, Genes, ras, Genetic Therapy, Gene Expression Regulation, Neoplastic, RNA, Catalytic, Apoptosis, Gene Silencing, RNA Interference, RNA, Small Interfering, Vascular Endothelial Growth Factor A, Cell Line, Tumor, Cell Proliferation
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