FHOD3 promotes carcinogenesis by regulating RhoA/ROCK1/LIMK1 signaling pathway in medulloblastoma

Clin Transl Oncol. 2020 Dec;22(12):2312-2323. doi: 10.1007/s12094-020-02389-5. Epub 2020 May 23.

Abstract

Purpose: Medulloblastoma (MB) is a malignant brain disease in young children. The overall survival of MB patients is disappointing due to absence of effective therapeutics and this could be attributed to the lack of molecular mechanism underlying MB. FHOD3 was an important gene during cardio-genesis and was reported to promote cell migration in cancer. However, its role in MB is not clear to date.

Methods: RT-qPCR and IHC analysis were used to determine expression of FHOD3. Survival curve was drawn by K-M analysis. FHOD3 was knocked down by RNAi technology. The effects of FHOD3 on medulloblastoma cells were determined by CCK-8 assay, colony formation assay, transwell assay and FACs analysis.

Results: FHOD3 expression increased by 1.5 fold in tumor tissues compared to the control and IHC analysis further confirmed strong expression of FHOD3 in medulloblastoma tissues. Then higher FHOD3 expression was associated with shorter survival time in MB patients (13.0 months versus 43.8 months). In medulloblastoma cells such as Daoy and D283med, FHOD3 also displayed abundant expression. When FHOD3 was knocked down, the ability of cell proliferation and colony formation was reduced over greatly. The capability of cell migration and invasion was also inhibited significantly. However, cell apoptotic rate increased significantly reversely. Mechanistically, the phosphorylation level of RhoA, ROCK1, and LIMK1 was decreased when FHOD3 was knocked down but increased reversely when FHOD3 was over-expressed in Daoy cells.

Conclusions: FHOD3 was associated with overall survival time in medulloblastoma patients and was essential to cell proliferation, growth and survival in medulloblastoma and might regulates activation of RhoA/ROCK1/LIMK1 signaling pathway.

Keywords: FHOD3; Medulloblastoma; ROCK1; RhoA.

MeSH terms

  • Apoptosis
  • Cell Cycle
  • Cell Line, Tumor
  • Cell Movement
  • Cell Proliferation
  • Cerebellar Neoplasms / metabolism*
  • Cerebellar Neoplasms / mortality
  • Child, Preschool
  • Female
  • Formins / genetics
  • Formins / metabolism*
  • Gene Knockdown Techniques
  • Humans
  • Kaplan-Meier Estimate
  • Lim Kinases / metabolism*
  • Male
  • Medulloblastoma / metabolism*
  • Medulloblastoma / mortality
  • Medulloblastoma / pathology
  • Neoplasm Invasiveness
  • Neoplasm Proteins / metabolism
  • Phosphorylation
  • Signal Transduction
  • Tumor Stem Cell Assay
  • rho-Associated Kinases / metabolism*
  • rhoA GTP-Binding Protein / metabolism*

Substances

  • FHOD3 protein, human
  • Formins
  • Neoplasm Proteins
  • RHOA protein, human
  • LIMK1 protein, human
  • Lim Kinases
  • ROCK1 protein, human
  • rho-Associated Kinases
  • rhoA GTP-Binding Protein