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The investigation of ceranib-2 on apoptosis and drug interaction with carboplatin in human non small cell lung cancer cells in vitro

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Abstract

Ceramide is found to be involved in inhibition of cell division and induction of apoptosis in certain tumour cells. Ceranib-2 is an agent that increases ceramide levels by inhibiting ceramidase in cancer cells. Therefore, we aimed to investigate the effects of ceranib-2 on cell survival, apoptosis and interaction with carboplatin in human non-small cell lung cancer cells. The cytotoxic effect of ceranib-2 (1–100 µM) was determined by MTT assay in human lung adenocarcinoma (A549) and large cell lung carcinoma (H460) cells. Carboplatin (1–100 µM) and lung bronchial epithelial cells (BEAS-2B) were used as positive controls. Morphological and ultrastructural changes were analysed by light microscope and TEM. Apoptotic/necrotic cell death and acid ceramidase activity were analysed by ELISA. Combination effects of ceranib-2 and carboplatin were investigated by MTT. The expression levels of CASP3, CASP9, BAX and BCL-2 were examined by qRT-PCR. The IC50 of ceranib-2 was determined as 22 μM in A549 cells and 8 μM in H460 cells for 24 h. Morphological changes and induction of DNA fragmentation have revealed apoptotic effects of ceranib-2 in both cell lines. Ceranib-2 and carboplatin has shown synergism in combined treatment at 10 and 25 μM doses in H460 cells for 24 h. Ceranib-2 inhibited acid ceramidase activity by 44% at 25 µM in H460 cells. Finally, CASP3, CASP9 and BAX expressions were increased while BCL-2 expression was reduced in both cells. Our results obtained some preliminary results about the cytotoxic and apoptotic effects of ceranib-2 for the first time in NSCLC cell lines.

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Acknowledgements

This publications was based completely on M.Sc. thesis of the first author, Merve YILDIZ.

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Authors and Affiliations

  1. Graduate School of Natural and Applied Sciences, Eskisehir Osmangazi University, Eskisehir, Turkey

    Merve Yildiz-Ozer

  2. Department of Biology, Faculty of Arts and Sciences, Eskisehir Osmangazi University, Eskisehir, Turkey

    Pinar Oztopcu-Vatan

  3. Department of Health Programme, Open Faculty, Anadolu University, Eskisehir, Turkey

    Gokhan Kus

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  1. Merve Yildiz-Ozer
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  2. Pinar Oztopcu-Vatan
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  3. Gokhan Kus
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Correspondence to Pinar Oztopcu-Vatan.

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This study was supported by TUBITAK (The Scientific and Technical Research Council of Turkey), (Project Number: 214Z159).

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Yildiz-Ozer, M., Oztopcu-Vatan, P. & Kus, G. The investigation of ceranib-2 on apoptosis and drug interaction with carboplatin in human non small cell lung cancer cells in vitro. Cytotechnology 70, 387–396 (2018). https://doi.org/10.1007/s10616-017-0154-8

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