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Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
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The gene encoding the main signaling molecules of the Wnt signaling pathways (the Wnt proteins) was discovered almost four decades ago by Nüsslein-Volhard and Wieschaus. They identified and originally named the gene "wingless" (wg) after a phenotype discovered during their landmark genetic screen in Drosophila for body pattern defects. At around the same time, another researcher named Harold Varmus found that a murine tumor virus activates the mammalian wg homolog, Int-1, which...
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Wnt is a zygotic effect gene that is expressed during very early embryonic development. It regulates various processes in animals starting from early development through the adult stage, such as organogenesis in the embryo and maintenance of neuronal and blood stem cells. Wnt proteins can induce a wide variety of intracellular pathways depending upon the specific abilities of different Wnt ligands to form a complex with shared and cognate receptors in the presence of different co-receptors. The...
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The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
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Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
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Updated: Mar 29, 2026

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Functional siRNA Screen Links Ras/MAPK and Wnt Pathway to EV Secretion in HCT-116 Colorectal Cancer Cells.

Sophie Marie Pätzold1, Julia Christina Gross2

  • 1Hematology and Oncology, University Medical Center Goettingen, 37075 Goettingen, Germany.

Diseases (Basel, Switzerland)
|March 27, 2026
PubMed
Summary

Investigating cancer-related genes in colorectal cancer cells revealed that most gene knockdowns increased extracellular vesicle (EV) secretion. Specific genes like KRAS and BRAF significantly impacted EV levels and characteristics.

Keywords:
EV secretionHCT-116-cellsRas/Raf/MAPK pathwayWnt pathwayexosomesmicrovesiclesoncogenesiRNA-mediated knockdowntumor suppressor gene

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Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • Extracellular vesicles (EVs) are crucial in tumor progression and cell communication.
  • The specific roles of cancer-related genes in EV secretion are not fully understood.

Purpose of the Study:

  • To investigate the impact of frequently altered cancer genes on EV secretion in colorectal cancer.
  • To identify specific genes influencing EV release and characteristics.

Main Methods:

  • Utilized siRNA-based loss-of-function screening in HCT-116 colorectal cancer cells.
  • Targeted 30 frequently altered (proto-)oncogenes and tumor suppressor genes.
  • Isolated and characterized EVs using ultracentrifugation, nanoparticle tracking analysis, and marker expression.

Main Results:

  • Most gene knockdowns led to increased EV secretion.
  • Silencing KRAS and BRAF significantly elevated EV levels, protein content, and particle size.
  • CTNNB1 and CDH1 showed distinct effects on different EV fractions.

Conclusions:

  • Components of Ras/Raf/MAPK and Wnt signaling pathways regulate EV secretion in colorectal cancer.
  • Gene-specific alterations influence EV release and characteristics, impacting intercellular communication in cancer.