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<p>Activating mutations in the KRAS oncogene, particularly the G12V substitution, are key drivers of tumorigenesis and therapeutic resistance across multiple cancer types. However, direct pharmacological inhibition of mutant KRAS has remained a major clinical challenge due to its structural and biochemical properties. In this study, we employed a chemically modified locked nucleic acid (LNA) GapmeR antisense oligonucleotide specifically designed to target the KRAS G12V transcript. The SW480 cell line was used as a representative in vitro model. Assays conducted were Quantitative RT-PCR, luciferase reporter assay using a KRAS representative construct, MTT assay, wound-healing assay, Annexin V-FITC apoptosis assay, and cell cycle distribution. RT-qPCR confirmed a marked reduction in KRAS mRNA levels. Luciferase reporter assay provided further evidence of GapmeR-mediated suppression at the transcript level. MTT revealed a robust cytotoxic effect at 100 nM, sufficient to induce significant cancer cell death while sparing normal cells. Scratch assay demonstrated increased scratch area, along with morphological changes. Apoptosis assay revealed a pronounced induction of late apoptosis and necrosis. Cell cycle analysis indicated disruption of the cell cycle. Our findings establish evidence that transcript level targeting of KRAS G12V using GapmeRs is associated with functional alterations in SW480 cells, warranting further validation in additional cancer models and in vivo studies.</p>

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LNA GapmeR silencing of KRAS G12V impairs growth and function in SW480 cells

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • Joudi Feras Khudeir,
  • Abdelaziz Tlili

摘要

Activating mutations in the KRAS oncogene, particularly the G12V substitution, are key drivers of tumorigenesis and therapeutic resistance across multiple cancer types. However, direct pharmacological inhibition of mutant KRAS has remained a major clinical challenge due to its structural and biochemical properties. In this study, we employed a chemically modified locked nucleic acid (LNA) GapmeR antisense oligonucleotide specifically designed to target the KRAS G12V transcript. The SW480 cell line was used as a representative in vitro model. Assays conducted were Quantitative RT-PCR, luciferase reporter assay using a KRAS representative construct, MTT assay, wound-healing assay, Annexin V-FITC apoptosis assay, and cell cycle distribution. RT-qPCR confirmed a marked reduction in KRAS mRNA levels. Luciferase reporter assay provided further evidence of GapmeR-mediated suppression at the transcript level. MTT revealed a robust cytotoxic effect at 100 nM, sufficient to induce significant cancer cell death while sparing normal cells. Scratch assay demonstrated increased scratch area, along with morphological changes. Apoptosis assay revealed a pronounced induction of late apoptosis and necrosis. Cell cycle analysis indicated disruption of the cell cycle. Our findings establish evidence that transcript level targeting of KRAS G12V using GapmeRs is associated with functional alterations in SW480 cells, warranting further validation in additional cancer models and in vivo studies.