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JS-K activates G2/M checkpoints through the DNA damage response and induces autophagy via CAMKKβ/AMPKα/mTOR pathway

Yuwan Zhao1, Shanhong Lin1, Wenfeng Zeng1

  • 1Laboratory of Urology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, China.

Journal of Cancer
|January 3, 2024
PubMed

Insights

JS-K, a nitric oxide donor, effectively inhibits bladder cancer cell growth by inducing DNA damage and cell cycle arrest. It also triggers autophagy via the CAMKKβ/AMPKα/mTOR pathway, offering a potential therapeutic strategy.

Area of Science:

  • Oncology
  • Molecular Biology
  • Biochemistry

Background:

  • Bladder cancer (BCa) remains a significant health concern, necessitating novel therapeutic agents.
  • Nitric oxide (NO) donor prodrugs are being explored for their anti-cancer properties.
  • Understanding the mechanisms of action, including DNA damage and autophagy induction, is crucial for drug development.

Purpose of the Study:

  • To investigate the effects of JS-K, a nitric oxide donor prodrug, on DNA damage and autophagy in bladder cancer cells.
  • To elucidate the underlying mechanisms of JS-K's anti-cancer activity.
  • To explore the potential of JS-K as a therapeutic agent for bladder cancer.

Main Methods:

  • Cell viability assays, morphology observation, and colony formation assays were used to assess JS-K's effect on BCa cell growth.
  • Western blot analysis was employed to detect DNA damage and repair proteins (γH2AX, PCNA, RAD51), cell cycle regulators, and autophagy-related proteins.
  • Flow cytometry was utilized to analyze cell cycle phase distribution and calcium levels.

Main Results:

  • Low concentrations of JS-K significantly inhibited BCa cell proliferation and colony formation without affecting normal cells.
  • JS-K induced DNA damage by increasing γH2AX and decreasing PCNA and RAD51 levels, while also causing G2/M phase arrest.
  • JS-K upregulated DNA damage checkpoint proteins (p-ATM, p-ATR, p-Chk1, p-Chk2, p-Cdc2) and downregulated Cyclin B1, leading to G2/M arrest.
  • JS-K induced autophagy in BCa cells, mediated by the CAMKKβ/AMPKα/mTOR pathway, and caused calcium overload.

Conclusions:

  • JS-K demonstrates selective anti-cancer effects on bladder cancer cells by inducing DNA damage and G2/M phase arrest.
  • The observed autophagy induction via the CAMKKβ/AMPKα/mTOR pathway is a key mechanism contributing to JS-K's anti-tumor activity.
  • JS-K holds promise as a potential therapeutic agent for bladder cancer, warranting further investigation.
Keywords:
AutophagyBladder cancerCell arrestDNA damageNO prodrug

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