Verbascoside potentiates the effect of tyrosine kinase inhibitors on the induction of apoptosis and oxidative stress via the Abl-mediated MAPK signalling pathway in chronic myeloid leukaemia

Abstract

Verbascoside (Verb) may exhibit potential antitumour activities in leukaemia. The present study investigated the effect of Verb, in combination with imatinib (IM), dasatinib (Das), lipopolysaccharide (LPS) and TNF, on cell survival, Abl expression, apoptosis, oxidative stress and the MAPK pathway in chronic myeloid leukaemia (CML) cells. Cell viability was determined using the WST-8 assay in K562 and R-K562 cells treated with Verb and/or IM, Das, LPS and TNF. Apoptosis and DNA damage in CML cells was detected by caspase-3 and comet analysis. The protein levels of Abl (Phospho-Tyr412), and total/phosphorylated p38, JNK and ERK in CML cells were analysed using a Colorimetric Cell-Based Assay. Oxidative stress was examined using total antioxidant and oxidant status assays. Treatment with Verb and/or tyrosine kinase inhibitors (TKIs), LPS and TNF resulted in a significant decrease in the Tyr-412 phosphorylation of Abl in K562 and R-K562 cells. In addition, cotreatment with Verb and IM or Das additively induced apoptosis by activating caspase-3 levels in both cell lines. Activation of p38 and JNK can result in growth arrest and cell death, whereas ERK stimulation results in cell division and differentiation. The present study demonstrated that cotreatment with Verb and TKIs suppressed phosphorylated-ERK1/2, whereas the levels of phosphorylated-p-38 and phosphorylated-JNK were significantly elevated by Verb and/or IM, Das, LPS and TNF, thus suggesting that Abl and Src inhibition could be involved in the effects of Verb on MAPK signalling in R-K562 cells. Furthermore, Verb elevated reactive oxygen species levels additively with TKIs in both cell lines by increasing the oxidant capacity and decreasing the antioxidant capacity. In conclusion, anti-leukemic mechanisms of Verb may be mediated by Abl protein and regulation of its downstream p38-MAPK/JNK pathway, caspase-3 and oxidative stress in CML cells.

FL118 Is a Potent Therapeutic Agent against Chronic Myeloid Leukemia Resistant to BCR-ABL Inhibitors through Targeting RNA Helicase DDX5

“…We first investigated the effects of imatinib on proliferation, survival, and DDX5 expression in K562 cells. With reference to previous reports [60,61], we chose two concentrations (1 and 2 µM) of imatinib to test their effects. However, there was no significant difference in the suppressive effects of 1 µM and 2 µM imatinib on them (Figure 1).…”

Calcium influx, oxidative stress, and apoptosis induced by TRPV1 in chronic myeloid leukemia cells: Synergistic effects with imatinib

“…On the other hand, the drug-induced oxidative stress exaggeration in leukemic cells could intensify the genotoxic effects of conventional therapy, facilitating the activation of programmed cell death ( Yang et al, 2018 ). In fact, recent findings showed that the stimulation of oxidative stress in CML, using compounds such as tyrosol derivatives, verbascoside, and zerumbone, promotes mitochondrial membrane potential dissipation, caspase activation, and apoptosis ( Rajan et al, 2015 ; Abdelkafi-Koubaa et al, 2022 ; Akgun-Cagliyan et al, 2022 ). Therefore, the strategy of deliberately increasing ROS to cytotoxic levels for leukemia cells seems to be very appealing, and our data are in agreement.…”

The Bone Marrow Immune Microenvironment in CML: Treatment Responses, Treatment-Free Remission, and Therapeutic Vulnerabilities

“…Plant-derived verbascoside can block CD44 dimerisation [51], and it was shown to induce apoptosis in a model of CML, possibly by mediating p38-MAPK/JNK and Caspase-3 signalling, although the role of CD44 was not explored here [52]. CD44 is highly expressed within normal, CML and AML CD34 + CD38 + and CD34 + CD38 − BM cells [53•], likely making it unsuitable as a CML LSC biomarker and a cancer-specific therapeutic target, but downstream signalling may be targetable.…”