Int J Stem Cells.  2018 Nov;11(2):227-234. 10.15283/ijsc18053.

The Pharmacological Inhibition of ERK5 Enhances Apoptosis in Acute Myeloid Leukemia Cells

• Kang C
• Kim JS
• Kim CY
• Kim EY
• Chung HM

• KMID: 2429555
• DOI: http://doi.org/10.15283/ijsc18053

Abstract

Acute myeloid leukemia (AML) is a fatal hematological malignancy which is resistant to a variety of chemotherapy drugs. Extracellular signal-regulated kinase 5 (ERK5) plays a novel role in chemoresistance in some cancer cells and this pathway is a central mediator of cell survival and apoptotic regulation. The aim of this study was to investigate the effect of ERK5 inhibitor, XMD8-92, on proliferation and apoptosis in AML cell lines. Findings showed that XMD8-92 inhibited the activation of ERK5 by G-CSF and decreased the expression of c-Myc and Cyclin D1. The treatment of XMD8-92 reduced the phosphorylation of ERK5 leading to a distinct inhibition of cell proliferation and increased apoptosis in Kasumi-1 and HL-60 cells. Taken together, our study suggests that the inhibition of ERK5 by XMD8-92 can trigger apoptosis and inhibit proliferation in AMLs. Therefore, the inhibition of ERK5 may be an effective adjuvant in AML chemotherapy.

Keyword

ERK5; XMD8-92; Cell cycle; Apoptosis; Acute myeloid leukemia

MeSH Terms

Apoptosis*
Cell Cycle
Cell Line
Cell Proliferation
Cell Survival
Cyclin D1
Drug Therapy
Granulocyte Colony-Stimulating Factor
Hematologic Neoplasms
HL-60 Cells
Humans
Leukemia, Myeloid, Acute*
Mitogen-Activated Protein Kinase 7
Phosphorylation
Cyclin D1
Granulocyte Colony-Stimulating Factor
Mitogen-Activated Protein Kinase 7

Figure

• 

  Fig. 1 Granulocyte colony-stimulating factor (G-CSF) induces the activation of ERK5 in AML cells. (A) The expression levels of ERK5 and ERK1/2 in AML cell lines (Kasumi-1, HL-60, MV4.11) by western blot analysis. (B) The phosphorylation of ERK5 and ERK1/2 in Kasumi-1 cells with the treatment of 10 ng/ml G-CSF.

• 

  Fig. 2 G-CSF-mediated ERK5 activation induces the proliferation of AML cells. (A, D) XMD8-92 suppressed the G-CSF-stimulated phosphorylation of ERK5 but not affected ERK1/2 in Kasumi-1 (A) and HL-60 (D) cells. (B, E) PD184352, as an inhibitor of ERK1/2, was not inhibited the G-CSF-stimulated activation of ERK5 in Kasumi-1 (B) and HL-60 (E) cells. (C, F) The cell proliferation induced by with or without G-CSF was suppressed by the treatment of XMD8-92 in Kasumi-1 (C) and HL-60 (F) cells, respectively. Data were obtained from three replicate experiments for the cell counting kit-8 assay. Data represent mean±SD. **p<0.01, ***p<0.005 indicates a significant difference compared to control.

• 

  Fig. 3 The inhibition of ERK5 by XMD8-92 induces apoptosis in AML cells. (A, B) The Kasumi-1 (A) and HL-60 (B) cells with or without treatments of G-CSF, XMD8-92, and PD184352 were stained with propidium iodide (PI) and then analyzed by flow cytometry. (C and D) The Kasumi-1 (C) and HL-60 (D) cells with or without treatments of G-CSF, XMD8-92, and PD184352 were stained with PE Annexin V and analyzed by flow cytometry.

• 

  Fig. 4 The inhibition of ERK5 by XMD-8-92 regulates on cell cycle- and apoptosis-related genes in AML cells. (A, B) The protein (A) and mRNA (B) levels of c-Myc decreased in Kasumi-1 cells through the inhibition of ERK5. (C and D) Analysis of the cell cycle- and apoptosis-related genes in Kasumi-1 (C) and HL-60 (D) cells by qRT-PCR. Relative expression of genes was calculated relative to GAPDH. Data represent mean±SD of 3 independent experiments. *p<0.05, **p<0.01, ***p<0.005 indicates a significant difference between samples.

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