Ginsenoside 24-OH-PD from red ginseng inhibits acute T-lymphocytic leukaemia by activating the mitochondrial pathway

PLoS One. 2023 May 19;18(5):e0285966. doi: 10.1371/journal.pone.0285966. eCollection 2023.

Abstract

Ginsenoside 24-hydroxy-ginsengdiol (24-OH-PD), extracted from red ginseng, is a novel diol-type ginsenoside, strongly inhibits the growth of human T-cell acute lymphoblastic leukaemia (T-ALL) CCRF-CEM cells. Our research aimed at investigating the mechanism underlying this inhibition. Cell viability was determined using the cell counting kit-8 (CCK-8) assay, and NOD/SCID mice bearing CCRF-CEM cells were used to verify the therapeutic effect of 24-OH-PD on T-ALL in vivo. We equally analysed pathways related to 24-OH-PD in CCRF-CEM cells using RNA-Seq analysis. Cell apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), and mitochondrial permeability transition pore (mPTP) levels were detected by flow cytometry. The activity of caspase3 and caspase9 was detected by enzyme activity detection kits. The expression levels of apoptosis-related proteins and mRNA were determined through western blotting and quantitative reverse-transcription PCR assays (qRT-PCR). CCK-8 assay and animal xenograft experiments confirmed that 24-OH-PD significantly inhibited T-ALL in a dose-dependent manner, both in vivo and in vitro. RNA-Seq results suggest that mitochondria-mediated apoptosis pathway plays an important role in this process. Furthermore, intracellular ROS levels increased, mPTP opened, and ΔΨm decreased following 24-OH-PD treatment. Pretreatment with the antioxidant, NAC, reversed the effects of 24-OH-PD on apoptosis and ROS generation. Moreover, 24-OH-PD treatment increased the expression of Bax and caspase family members, thereby releasing cytochrome c (Cytc) and inducing apoptosis. Our findings showed that, 24-OH-PD induces apoptosis in CCRF-CEM cells by activating the mitochondrial-dependent apoptosis pathway through ROS accumulation. This inhibitory effect implies that 24-OH-PD could be further developed as treatment of T-ALL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Apoptosis Regulatory Proteins / metabolism
  • Ginsenosides* / metabolism
  • Ginsenosides* / pharmacology
  • Humans
  • Membrane Potential, Mitochondrial
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Mitochondria / metabolism
  • Panax* / metabolism
  • Precursor T-Cell Lymphoblastic Leukemia-Lymphoma* / metabolism
  • Reactive Oxygen Species / metabolism

Substances

  • Ginsenosides
  • Reactive Oxygen Species
  • Apoptosis Regulatory Proteins

Grants and funding

This work was supported by the China PostDoctoral Science Foundation (Grant No. 2018M641678), National Natural Science Foundation for Young Scholars of China (Grant No. 82104667), Youth Fund of Basic Research Program of Shanxi Province (Grant No. 2021030212456, 201801D221272), and Academic Project of the Shanxi Academy of Traditional Chinese Medicine (Grant No. 202101). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.