α-Tomatine, a novel early-stage autophagy inhibitor, inhibits autophagy to enhance apoptosis via Beclin-1 in Skov3 cells

Fitoterapia. 2021 Jul:152:104911. doi: 10.1016/j.fitote.2021.104911. Epub 2021 Apr 24.

Abstract

Targeting the autophagy process is considered to be a promising new strategy for drug treatment of ovarian cancer. α-Tomatine, a steroidal alkaloid extracted, is mainly isolated from leaves, roots and immature green tomatoes. α-Tomatine has biological activities such as anticancer, antioxidative and anti-inflammatory. The study aimed to explore the effects of α-tomatine on proliferation, apoptosis and autophagy and the underlying mechanisms in ovarian cancer Skov3 cells. After treatment with different concentrations of α-tomatine (0, 0.75, 1 and 1.5 μM) in Skov3 cells for 24 h, proliferation was determined by the CCK-8 assay, and apoptosis was detected by flow cytometric analysis. Autophagy in cells was determined by the number of fluorescent spots using confocal fluorescence microscopy after mRFP-GFP-LC3 transfection. The relationship between autophagy and apoptosis was proved by Beclin-1 overexpression. The protein expression levels were tested by western blotting. The results demonstrated that α-tomatine effectively repressed proliferation, exerted a proapoptotic effect and inhibited early-stage autophagy in Skov3 cells in a dose- and time-dependent manner. Additionally, Beclin-1 overexpression significantly suppressed α-tomatine-treated apoptosis in Skov3 cells, indicating that α-tomatine inhibits autophagy to induce apoptosis. We also found α-tomatine inhibited the protein expression levels of PI3K/Akt/mTOR signaling pathway. However, the autophagy inhibition of α-tomatine could be reversed obviously by Beclin-1 overexpression. Taken together, α-tomatine inhibited autophagy through Beclin-1. Our study suggests that α-tomatine, as a novel early-stage autophagy inhibitor, might be a potential drug for further treatment of ovarian cancer by inhibiting proliferation and promoting apoptosis.

Keywords: Apoptosis; Autophagy inhibitor; Beclin-1; Ovarian cancer; α-Tomatine.

MeSH terms

  • Apoptosis / drug effects*
  • Autophagy / drug effects*
  • Beclin-1 / metabolism*
  • Cell Line, Tumor
  • Female
  • Humans
  • Molecular Structure
  • Ovarian Neoplasms / drug therapy
  • Signal Transduction
  • Solanum lycopersicum / chemistry
  • Tomatine / analogs & derivatives*
  • Tomatine / pharmacology

Substances

  • BECN1 protein, human
  • Beclin-1
  • alpha-tomatine
  • Tomatine