Jolkinolide B attenuates laryngeal cancer cell growth and induces apoptosis via PTEN/PI3K/Akt signaling pathway

In Vitro Cell Dev Biol Anim. 2021 Sep;57(8):786-794. doi: 10.1007/s11626-021-00612-3. Epub 2021 Oct 25.

Abstract

Jolkinolide B (JB) is a bioactive diterpenoid, isolated from the root of Euphorbia fischeriana Steud, and has been reported to have anti-tumor and anti-inflammation function by regulation of cell migration, invasion, apoptosis, and cell cycle. We aimed to evaluate the effect of JB on laryngeal cancer cells. Human normal larynx epithelial (HBE) cells and cancer cell lines TU212, TU177, and Hep-2 were cultured; MTT assay was used to assess cell proliferation. LY294002 (a PI3K/Akt inhibitor) and IGF-1 (a PI3K/Akt activator) were employed to investigate the expression of PI3K/Akt pathway. Cell migration and invasion activities were detected by scratch wound healing and transwell assay, respectively. Flow cytometry assay was used to assess cell apoptosis. The expression levels of proteins were assessed by immunofluorescence and Western blotting assay. JB inhibited TU212, TU177, and Hep-2 cell viability with an IC50 value of 54.57 ± 0.53 μg/mL, 44.82 ± 0.32 μg/mL, and 49.63 ± 0.47 μg/mL, respectively. Compared with control group, the proliferation, migration, and invasion of cells significantly decreased after JB and LY294002 treatment, while cell apoptosis increased. In IGF-1 group, the results were opposite compared to the JB and LY294002 groups. Western blotting results showed that JB and LY294002 treatment significantly inhibited the levels of Bcl-2, p-PI3K, and p-Akt while the levels of Bax, cleaved caspase-3, and PTEN protein significantly increased. Our study suggested that JB exhibits an inhibition effect on laryngeal cancer cell growth in vitro.

Keywords: Cell proliferation; Cleaved caspase-3; Jolkinolide B; Laryngeal cancer; PTEN.

MeSH terms

  • Antineoplastic Agents, Phytogenic / therapeutic use*
  • Apoptosis / drug effects*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Chromones / pharmacology
  • Diterpenes / therapeutic use*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Humans
  • Hylobatidae
  • Insulin-Like Growth Factor I / pharmacology
  • Laryngeal Neoplasms / drug therapy*
  • Morpholines / pharmacology
  • PTEN Phosphohydrolase / metabolism*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction / drug effects*

Substances

  • Antineoplastic Agents, Phytogenic
  • Chromones
  • Diterpenes
  • Morpholines
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • jolkinolide B
  • Insulin-Like Growth Factor I
  • Proto-Oncogene Proteins c-akt
  • PTEN Phosphohydrolase
  • PTEN protein, human

Supplementary concepts

  • Nomascus concolor