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CYLD destabilizes NoxO1 protein by promoting ubiquitination and regulates prostate cancer progression

Saba Haq1, Neha Sarodaya2, Janardhan Keshav Karapurkar2

  • 1Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, 04763, South Korea.

Cancer Letters
|November 7, 2021
PubMed

Insights

Cylindromatosis (CYLD) deubiquitinating enzyme destabilizes Nox organizer 1 (NoxO1) protein, suppressing excessive reactive oxygen species (ROS) and cancer cell growth. CYLD acts as a tumor suppressor, offering a potential therapeutic target for cancer treatment.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • NADPH oxidase (Nox) enzymes generate reactive oxygen species (ROS), crucial in cell signaling and various diseases, including cancer.
  • Maintaining cellular ROS balance requires strict regulation of Nox enzyme activity.
  • Dysregulated ROS production is implicated in cancer progression.

Purpose of the Study:

  • To identify deubiquitinating enzymes (DUBs) that regulate Nox organizer 1 (NoxO1) protein expression.
  • To investigate the role of cylindromatosis (CYLD) in NoxO1 regulation and its impact on cancer.
  • To explore CYLD as a potential therapeutic target in cancer treatment.

Main Methods:

  • Genome-wide screening using a CRISPR/Cas9-mediated DUB-knockout library to identify regulators of NoxO1.
  • Co-immunoprecipitation to confirm CYLD as a binding partner of NoxO1.
  • Western blotting and ubiquitination assays to assess NoxO1 protein levels and stability.
  • In vitro cell proliferation, migration, colony formation, and invasion assays using CYLD-depleted cells.
  • In vivo tumor growth studies in xenografted mice using CYLD-depleted cancer cells.

Main Results:

  • CYLD was identified as a regulator of NoxO1 protein expression.
  • Overexpression of CYLD led to increased ubiquitination and reduced half-life of NoxO1 protein.
  • CYLD-mediated destabilization of NoxO1 suppressed excessive ROS generation.
  • CRISPR/Cas9-mediated knockout of CYLD in PC-3 cells enhanced cell proliferation, migration, colony formation, and invasion.
  • Tumor growth in xenografted mice was increased when using CYLD-depleted cancer cells, with larger tumor weight and volume.

Conclusions:

  • CYLD functions as a destabilizer of NoxO1 protein, thereby controlling ROS levels.
  • CYLD exhibits tumor-suppressive properties by inhibiting cancer cell proliferation, migration, and invasion.
  • CYLD represents a promising therapeutic target for developing novel cancer treatments.
Keywords:
CRISPR/Cas9Colon cancerDeubiquitinationProteasomal degradationProteolysisROS

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