Synergistic killing effects of homoharringtonine and arsenic trioxide on acute myeloid leukemia stem cells and the underlying mechanisms

Abstract

Background At present, it is generally believed that leukemia stem cells are the source of AML, so the killing of leukemia stem cells has become important. Previous studies have suggested that HHT combined with ATO can synergistically kill U937 cells, and HHT has also demonstrated the ability to kill leukemia stem cells. We evaluated whether HHT combined with ATO can systematically kill leukemia stem cells (LSCs) and explored the synergistic effect and molecular mechanism. Methods CCK-8 was used to detect cell viability. The changes of cell cycle (PI staining), apoptosis (Annexin V/PI) and surface markers (CD34, CD38, CD96, CD45) were detected by flow cytometry. The cells of CD34+ primary leukemia and CD38- KG-1, and TF-1 were separated by flow cytometry. High-throughput mRNA sequencing was used to analysis mRNA level changes after the application of the two drugs. Western blot was used to verify the changes of pathway protein expression. NRG mice were used as the receptor of xenograft model. Histological H&E staining assess the invaded ability of leukemia cells, and laser scanning confocal microscopy evaluated the molecule markers change. Results HHT and ATO synergistically killed KG-1 (CD34 + /CD96 + /CD38 + / − ) and Kasumi-1 (CD34 + /CD38 − ) cells. Their combination had a stronger effect of inducing apoptosis and blocking the cell cycle than HHT or ATO administrator alone, meanwhile significantly reducing the numbers of LSCs. Further, CD34 + CD38 − cells in KG-1, KG-1a, TF-1, and primary leukemia cells were more sensitive to HHT and ATO. High-throughput mRNA sequencing suggested that HHT alone could significantly upregulate molecules related to the Notch, P53, and NF-κB signaling pathways. When combined with ATO, HHT further upregulated P53, whereas HHT-induced NF-κB pathway activation was significantly suppressed. Western blot analysis verified the change of protein expression in the above pathways and further demonstrated that GSI, could eliminate these effects. In vivo, HHT combined with ATO significantly reduced the LSC burden, and weakened the expression of LSC markers. Conclusions This is the first evidence that HHT combined with arsenic can synergistically kill LSCs in vitro and in vivo, along with identification of the underlying mechanism, highlighting a potentially effective treatment strategy. Electronic supplementary material The online version of this article (10.1186/s13046-019-1295-8) contains supplementary material, which is available to authorized users.

Targeting the Human 80S Ribosome in Cancer: From Structure to Function and Drug Design for Innovative Adjuvant Therapeutic Strategies

“…The combination of HHT with arsenic trioxide was efficient to kill CD34 + CD38 − cells in the KG-1 and Kasumi-1 AML cell lines and in primary cells, suggesting that the treatment could affect Leukemic Stem Cells that maintain the disease and are responsible for relapse. Although, this assumption would need more in vivo experiments [115].…”

Correction to: Synergistic killing effects of homoharringtonine and arsenic trioxide on acute myeloid leukemia stem cells and the underlying mechanisms

“…In the publication of this article [1], there are two corrections: 1. The corresponding author Yuanzhong Chen’s email should be changed to chenyz@mail.fjmu.edu.cn; 2.…”

Tumor microenvironment: a prospective target of natural alkaloids for cancer treatment

“…The mechanism of its anti-myeloma may be the inhibition of AKT phosphorylation and several AKT target genes including NF-κB, XIAP, cIAP and Cyclin D1, the suppression of MCL1 protein synthesis, and the induction of apoptosis in chronic lymphocytic leukemia cells. HHT alone also significantly up-regulates the signal pathway of Notch, p53 and NF-κB [49][50][51]. Chronic Lymphocytic Leukemia (CLL) is related to T cell dysfunction.…”