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Research Article

Myrtus communis L. essential oil reduced colony formation and altered cell differentiation markers in lung cancer spheroids

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Pages 626-638 | Received 18 Mar 2023, Accepted 29 Jun 2023, Published online: 22 Aug 2023
 

Abstract

It was aimed to investigate the effect of Myrtus communis L. (MC) essential oil on cancer spheroids derived from A549 cells by analyzing cell viability and expression levels of prominent cancer stem cell and apoptosis markers. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide assay was per-formed to analyze cytotoxicity of MC essential oil for 24-h and 72-h. A549 cells were cultured in plates with low adherence to obtain spheroids. The effect of MC essential oil was assessed on the clonogenicity of the spheroids. Quantitative reverse-transcription polymerase chain reaction method was utilized to measure relative expressions of cluster of differentiation 44 (CD44), prominin-1 (PROM1), POU class 5 homeobox 1 (POU5F1), ATP binding cassette subfamily G member-2 (ABCG2), and caspase-3 (CASP3). Relative protein levels of ABCG2 and CASP3 were also evaluated with Western blot analysis. MC essential oil reduced cell viability of A549 cells after 24-h and 72-h. A reduction was observed in number of cancer spheroids after MC essential oil treatment for 7 days. After 24h of MC essential oil treatment, PROM1, POU5F1, ABCG2, and CASP3 gene expressions decreased while returned to basal levels after 72h. CD44 levels did not change upon MC essential oil treatment. ABCG2 protein level reduced after 24-h MC essential oil treatment and returned to its basal level after 72-h. CASP3 protein levels did not change at either time points. MC essential oil exerted its anti-tumorigenic effects onto lung cancer spheroids derived from A549 cells by reducing cell viability of cancer cells, inhibiting spheroid formation efficiency, and downregulating several cancer stem cell markers.

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