Harmine, a dual-specificity tyrosine phosphorylation-regulated kinase (DYRK) inhibitor induces caspase-mediated apoptosis in neuroblastoma

Abstract

BackgroundNeuroblastoma (NB) is an early childhood malignancy that arises from the developing sympathetic nervous system. Harmine is a tricyclic β-carboline alkaloid isolated from the harmal plant that exhibits both cytostatic and cytotoxic effects. Harmine is capable of blocking the activities of dual-specificity tyrosine phosphorylation-regulated kinase (DYRK) family proteins and mitogen-activated protein kinase. These kinases promote proliferation and inhibit apoptosis.MethodsFour human NB cell lines were used to study the effects of harmine treatment: SKNBE and KELLY (MYCN-amplified) as well as SKNAS and SKNFI (MYCN non-amplified). The anti-cancer properties of harmine were examined by RealTime-Glo MT cell viability assays, caspase activity assays, PARP cleavage using Western blot analysis, and flow cytometry-based Annexin V detection. A molecular interaction model of harmine bound to the DYRK2 family kinase was generated by computational docking using X-ray structures. NB tumors from human patients were profiled for DYRK mRNA expression patterns and clinical correlations using the R2 platform.ResultsThe IC50 values for harmine after 72 h treatment were 169.6, 170.8, and 791.7 μM for SKNBE, KELLY, and SKNFI, respectively. Exposure of these NB cell lines to 100 μM of harmine resulted in caspase-3/7 and caspase-9 activation as well as caspase-mediated PARP cleavage and Annexin V-positive stained cells, as early as 24 h after treatment, clearly suggesting apoptosis induction, especially in MYCN-amplified cell lines. Elevated DYRK2 mRNA levels correlated with poor prognosis in a large cohort of NB tumors.ConclusionHarmine is a known inhibitor of DYRK family kinases. It can induce apoptosis in NB cell lines, which led us to investigate the clinical correlations of DYRK family gene expression in NB tumors. The patient results support our hypothesis that DYRK inhibition by harmine and the subsequent triggering of caspase-mediated apoptosis might present a novel approach to NB therapy.Electronic supplementary materialThe online version of this article (10.1186/s12935-018-0574-3) contains supplementary material, which is available to authorized users.

Harmine induces anticancer activity in breast cancer cells via targeting TAZ

“…Previous studies have reported the antitumor effects of HM on hepatocellular carcinoma (39), gastric cancer (9) and neuroblastoma (40). In the present study, the anticancer activity of HM was demonstrated using MDA-MB-231 and MCF-7 cells in vitro and MCF-7 cell-derived xenograft tumor in vivo .…”

The DYRK Family of Kinases in Cancer: Molecular Functions and Therapeutic Opportunities

“…However, EGCG can potentially target many different intracellular pathways [ 177 ], making it difficult to assign particular effects to DYRK1A inhibition. Additionally, the ß-carboline alkaloid harmine selectively inhibits DYRK1A and—albeit, less efficiently—other members of the family [ 178 , 179 ], and it has been reported to have cytotoxic effects on cancer cell lines [ 66 , 85 , 180 , 181 ] and antitumor effects in vivo in glioma and in PDAC xenograft experiments [ 66 , 68 ], as well as synergistic effects with other chemotherapeutic agents [ 79 , 80 , 182 ]. However, the neurotoxic effects of harmine due to the targeting of monoamine oxidase A rule against its use in humans.…”

Updating dual-specificity tyrosine-phosphorylation-regulated kinase 2 (DYRK2): molecular basis, functions and role in diseases

“…For example, there is a positive correlation of DYRK2 expression with patient survival in hepatocellular, non-small cell lung, pulmonary adenocarcinoma, colorectal and bladder cancers, and non-Hodgkin's lymphoma [76,77,[80][81][82][83][84]. On the contrary, other results have revealed that DYRK2 expression in neuroblastoma inversely correlates with patient survival [85]. In contrast to the previously observed positive relation in breast cancer [43,86], Guo et al observed an inverse correlation between DYRK2 expression and survival in the same cancer subtype [66].…”

“…As a cell permeable inhibitor, it is commonly used to study DYRK family functionality [92]. It inhibits cell proliferation, migration, and invasion in several human cancer cell lines [85,93].…”