NEK6 Knockout Causes Defects in Mitochondrial Morphology and Respiration

Area of Science:

• Cell Biology
• Molecular Biology
• Cancer Research

Background:

• Nek kinases regulate cell cycle and mitochondrial homeostasis.
• Novel roles for Nek kinases in mitochondrial metabolism are emerging.
• NEK6’s role in mitochondrial function remained largely uncharacterized.

Purpose of the Study:

• To investigate the impact of NEK6 gene knockout on mitochondrial respiration and morphology in DU-145 prostate cancer cells.
• To elucidate the molecular mechanisms underlying NEK6-mediated mitochondrial regulation.

Main Methods:

• CRISPR-Cas9 gene editing to generate NEK6 knockout DU-145 cells.
• Transmission electron microscopy (TEM) for mitochondrial morphology analysis.
• Western blot and molecular assays for protein expression and mitochondrial function markers.

Main Results:

• NEK6 knockout cells showed reduced mitochondrial respiration and fragmented mitochondria with fewer cristae.
• Increased expression of anaerobic glycolysis markers (HK2, PFKP, LDHA) and decreased aerobic glycolysis marker (PDH).
• Altered mitochondrial dynamics (OPA1, DRP1), increased ER-mitochondria contact, and enhanced autophagy markers (LC3A/B, BECLIN1).

Conclusions:

• NEK6 plays a crucial role in maintaining mitochondrial homeostasis, including morphology and respiration.
• Loss of NEK6 promotes a shift towards anaerobic glycolysis and triggers mitochondrial degradation through autophagy.
• These findings suggest a novel function for NEK6 in regulating mitochondrial quality control in cancer cells.