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Function of a complex of p-Y42 RhoA GTPase and pyruvate kinase M2 in EGF signaling pathway in glioma cells

Yoon-Beom Lee1,2, Yohan Park1, Amir Hamza1,2

  • 1Department of Biochemistry, Hallym University, Chuncheon, Kangwon-do, Republic of Korea.

Journal of Neurochemistry
|August 26, 2024
PubMed

Insights

Epidermal growth factor (EGF) stimulates glioma cell proliferation. Phosphorylated RhoA (p-Y42 RhoA) and pyruvate kinase M2 (PKM2) interact to regulate PGK1, driving glioma progression.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Cell Signaling

Background:

  • Epidermal growth factor (EGF) is a known stimulant for glioma cancer cell proliferation.
  • RhoA protein levels and phosphorylation are altered by EGF in glioma cells.
  • Pyruvate kinase M2 (PKM2) is implicated in cancer cell metabolism and growth.

Purpose of the Study:

  • To elucidate the novel mechanism by which EGF-induced signaling pathways contribute to glioma progression.
  • To investigate the interaction between RhoA, PKM2, and downstream targets in glioma cells.
  • To identify key regulators of PGK1 expression in the context of EGF stimulation.

Main Methods:

  • In vitro binding assays to study protein interactions (GST-RhoA and PKM2).
  • Western blotting and immunoprecipitation to analyze protein levels, phosphorylation, and interactions.
  • siRNA-mediated knockdown to assess the functional role of RhoA.
  • Nuclear localization studies using EGF stimulation.
  • Reporter assays to evaluate gene promoter activity (PGK1).

Main Results:

  • EGF reduced RhoA protein but increased p-Y42 RhoA, PKM1, and PKM2 in LN18 glioma cells.
  • RhoA degradation involves ubiquitination by SCF1 and Smurf1.
  • p-Y42 RhoA specifically binds to PKM2, and PKM2 stabilizes RhoA.
  • EGF stimulation led to nuclear localization of RhoA, p-Y42 RhoA, and PKM2.
  • RhoA knockdown reduced PGK1 and MARK4 levels.
  • p-Y42 RhoA and PKM2 co-immunoprecipitated with β-catenin and YAP, which are associated with the PGK1 promoter.

Conclusions:

  • A novel mechanism is proposed where p-Y42 RhoA and PKM2, along with β-catenin and YAP, regulate PGK1 expression.
  • This pathway contributes to glioma progression upon EGF stimulation.
  • Targeting the p-Y42 RhoA/PKM2/β-catenin/YAP/PGK1 axis may offer therapeutic strategies for gliomas.
Keywords:
EGFPGK1PKM2gliomap‐Y42 RhoA

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