Compound kushen injection suppresses human acute myeloid leukaemia by regulating the Prdxs/ROS/Trx1 signalling pathway

Yanxia Jin,Dongdong Zhang,Li Liang,Yi Yang,Yunbao Pan,Tian Yang,Lu Ding,Honglei Tu,Hui Shen,Balu Wu,Hailing Liu,Qian Yang,Fuling Zhou,Yuxing Liang,Yongchang Wei,Tingting Huang

doi:10.1186/s13046-018-0948-3

Abstract

BackgroundThe increase in the levels of reactive oxygen species (ROS) in acute myeloid leukemia (AML) patients has been previously described; thus, it is important to regulate ROS levels in AML.MethodsFlow cytometry were used to assess the in vitro effect of compound kushen injection (CKI). Quantitative proteomics were used to analyse the mechanism. The AML patient-derived xenograft (PDX) model were used to evaluate the in vivo effect of CKI.ResultsWe found that intracellular ROS levels in AML cells were decreased, the antioxidant capacity were increased when treated with CKI. CKI inhibited the proliferation of AML cells and enhanced the cytotoxicity of AML cells, which has few toxic effects on haematopoietic stem cells (HSCs) and T cells. At the single-cell level, individual AML cells died gradually by CKI treatment on optofluidic chips. CKI promoted apoptosis and arrested cell cycle at G1/G0 phase in U937 cells. Furthermore, higher peroxiredoxin-3 (Prdx3) expression levels were identified in CKI-treated U937 cells through quantitative proteomics detection. Mechanically, the expression of Prdx3 and peroxiredoxin-2 (Prdx2) was up-regulated in CKI-treated AML cells, while thioredoxin 1 (Trx1) was reduced. Laser confocal microscopy showed that the proteins Prdx2 could be Interacted with Trx1 by CKI treatment. In vivo, the survival was longer and the disease was partially alleviated by decreased CD45+ immunophenotyping in peripheral blood in the CKI-treated group in the AML PDX model.ConclusionsAntioxidant CKI possess better clinical application against AML through the Prdxs/ROS/Trx1 signalling pathway.

Highlights

The increase in the levels of reactive oxygen species (ROS) in acute myeloid leukemia (AML) patients has been previously described; it is important to regulate ROS levels in AML
Antioxidant compound kushen injection (CKI) decreased intracellular ROS levels in AML cells After treatment with CKI, the intracellular ROS levels in THP-1, Molm-13, HL60, U937 and human AML cells were significantly reduced compared with controls (Fig. 1a)
The total antioxidant capacity (T-AOC) activity and GSH contents significantly increased in HL60 cells, Molm-13 cells and human AML cells, but there was no changed in THP-1 cells and had a slightly increase trendency in U937 cells

Summary

Introduction

The increase in the levels of reactive oxygen species (ROS) in acute myeloid leukemia (AML) patients has been previously described; it is important to regulate ROS levels in AML. Hole et al proposed that more than 60% of AML patients had sustained NADPH oxidase (NOX) activation, leading to high ROS accumulation and increased AML cell proliferation [2]. An increase in ROS levels is closely related to DNA oxidative damage and mutation in the development of AML [3]. Similar to our previous reports, the increased ROS levels in AML patient relapse induced higher promotor oncogene c-Jun activation domain-binding protein l (Jab1) and thioredoxin 1 (Trx1) expression; Jab binding to Trx affected Trx1 [6]. The higher ROS levels and Jab and Trx expression were significantly positively correlated with poor survival in AML patients, which promoted malignant proliferation in AML cells. Regulating the ROS signalling pathway will be a promising strategy for AML treatment

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