Fstl1 Promotes Glioma Growth Through the BMP4/Smad1/5/8 Signaling Pathway

Cell Physiol Biochem. 2017;44(4):1616-1628. doi: 10.1159/000485759. Epub 2017 Dec 6.

Abstract

Background: Gliomas result in the highest morbidity and mortality rates of intracranial primary central nervous system tumors because of their aggressive growth characteristics and high postoperative recurrence. They are characterized by genetic instability, intratumoral histopathological variability and unpredictable clinical behavior in patients. Proliferation is a key aspect of the clinical progression of malignant gliomas, complicating complete surgical resection and enabling tumor regrowth and further proliferation of the surviving tumor cells.

Methods: The expression of Fstl1 was detected by western blotting and qRT-PCR. We used cell proliferation and colony formation assays to measure proliferation. Then, flow cytometry was used to analyze cell cycle progression. The expression of Fstl1, p-Smad1/5/8 and p21 in GBM tissue sections was evaluated using immunohistochemical staining. Furthermore, we used coimmunoprecipitation (Co-IP) and immunoprecipitation to validate the relationship between Fstl1, BMP4 and BMPR2. Finally, we used orthotopic xenograft studies to measure the growth of tumors in vivo.

Results: We found that follistatin-like 1 (Fstl1) was upregulated in high-grade glioma specimens and that its levels correlated with poor prognosis. Fstl1 upregulation increased cell proliferation, colony formation and cell cycle progression, while its knockdown inhibited these processes. Moreover, Fstl1 interacted with bone morphogenetic protein (BMP) 4, but not BMP receptor (BMPR) 2, and competitively inhibited their association. Furthermore, Fstl1 overexpression suppressed the activation of the BMP4/Smad1/5/8 signaling pathway, while BMP4 overexpression reversed this effect.

Conclusion: Our study demonstrated that Fstl1 promoted glioma growth through the BMP4/Smad1/5/8 signaling pathway, and these findings suggest potential new glioblastoma treatment strategies.

Keywords: BMP4; Fstl1; Glioblastoma; Proliferation; Smad1/5/8 signaling pathway.

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 4 / metabolism*
  • Bone Morphogenetic Protein Receptors, Type II / metabolism
  • Brain Neoplasms / metabolism
  • Brain Neoplasms / mortality
  • Brain Neoplasms / pathology*
  • Cell Line, Tumor
  • Follistatin-Related Proteins / antagonists & inhibitors
  • Follistatin-Related Proteins / genetics
  • Follistatin-Related Proteins / metabolism*
  • Glioma / metabolism
  • Glioma / mortality
  • Glioma / pathology*
  • Humans
  • Immunoprecipitation
  • Kaplan-Meier Estimate
  • Mice
  • Phosphorylation
  • Protein Binding
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • S Phase Cell Cycle Checkpoints
  • Signal Transduction
  • Smad1 Protein / metabolism*
  • Smad5 Protein / metabolism*
  • Smad8 Protein / metabolism*
  • Transplantation, Heterologous

Substances

  • Bone Morphogenetic Protein 4
  • Follistatin-Related Proteins
  • RNA, Small Interfering
  • Smad1 Protein
  • Smad5 Protein
  • Smad8 Protein
  • FSTL1 protein, human
  • Bone Morphogenetic Protein Receptors, Type II