Origanum Sipyleum Methanol Extract in Combination with Ponatinib Shows Synergistic anti-Leukemic Activities on Chronic Myeloid Leukemia Cells

Abstract

Origanum sipyleum is used in folk medicine due to its anti-inflammatory, antimicrobial, and antioxidant properties. Ponatinib, an effective tyrosine kinase inhibitor in the treatment of chronic myeloid leukemia (CML), has severe side effects. Thus, we aimed to determine a novel herbal combination therapy that might not only increase the anti-leukemic efficacy but also reduce the dose of ponatinib in targeting CML cells. Origanum sipyleum was extracted with methanol (OSM), and secondary metabolites were determined by phytochemical screening tests. The cytotoxic effects of OSM on K562 cells were measured by WST-1 assay. Median-effect equation was used to analyze the combination of ponatinib and OSM (p-OSM). Apoptosis, proliferation, and cell-cycle were investigated by flow-cytometry. Cell-cycle-related gene expressions were evaluated by qRT-PCR. OSM that contains terpenoids, flavonoids, tannins, and anthracenes exhibited cytotoxic effects on K562 cells. The median-effect of p-OSM was found as synergistic; OSM reduced the ponatinib dose ∼5-fold. p-OSM elevated the apoptotic and anti-proliferative activity of ponatinib. Consistently, p-OSM blocked cell-cycle progression in G₀/G₁, S phases accompanied by regulations in TGFB2, ATR, PP2A, p18, CCND1, CCND2, and CCNA1 expressions. OSM enhanced the anti-leukemic activity of ponatinib synergistically via inducing apoptosis, suppressing proliferation, and cell-cycle. As a result, OSM might offer a potential strategy for treating patients with CML.

Acknowledgments

The Origanum sipyleum L. methanol extract was obtained from TÜBİTAK project no 110S289. We thank Ege University School of Foreign Languages and Lecturer Dr. Atiyye Hilal SENGENC for providing professional English editing for this manuscript.

Disclosure Statement

No potential conflict of interest was reported by the authors.

Authors’ Contributions

Conceptualization: Kayabasi C, Gunduz C. Design: Kayabasi C, Saydam G, Durmuskahya C, Kayalar H, Ozbilgin A, Biray Avci C, Gunduz C. Supervision/Consulting: Saydam G, Biray Avci C, Gunduz C. Data collection: Kayabasi C, Yilmaz Susluer S, Balci Okcanoglu T,Ozmen Yelken B, Mutlu Z, Goker Bagca B, Caliskan Kurt C, Kayalar H. Analysis: Kayabasi C, Yilmaz Susluer S, Balci Okcanoglu T, Ozmen Yelken B, Mutlu Z, Goker Bagca B, Caliskan Kurt C, Kayalar H, Gunduz C. Literature review for the study: Kayabasi C, Yilmaz Susluer S, Balci Okcanoglu T,Ozmen Yelken B, Mutlu Z, Goker Bagca B, Caliskan Kurt C, Saydam G, Durmuskahya C, Kayalar H, Ozbilgin A, Biray Avci C, Gunduz C. Writing the manuscript: Kayabasi C. Critical feedback: Saydam G, Durmuskahya C, Kayalar H, Ozbilgin A, Biray Avci C, Gunduz C. Final approval: Kayabasi C, Yilmaz Susluer S, Balci Okcanoglu T,Ozmen Yelken B, Mutlu Z, Goker Bagca B, Caliskan Kurt C, Saydam G, Durmuskahya C, Kayalar H, Ozbilgin A, Biray Avci C, Gunduz C.

Data Availability Statement

The data are available from the corresponding author upon reasonable request.

Ethics Approval and Consent to Participate

The local ethics committee ruled that no formal ethics approval was required in this particular case.

Funding

The author(s) reported there is no funding associated with the work featured in this article.