Multi‑drug‑resistance efflux in cisplatin‑naive and cisplatin‑exposed A2780 ovarian cancer cells responds differently to cell culture dimensionality

Koshkin,Vasilij; De Oliveira,Mariana Bleker; Peng,Chun; Ailles,Laurie E.; Liu,Geoffrey; Covens,Allan; Krylov,Sergey N.

doi:10.3892/mco.2021.2323

Multi‑drug‑resistance efflux in cisplatin‑naive and cisplatin‑exposed A2780 ovarian cancer cells responds differently to cell culture dimensionality

Authors:
- Vasilij Koshkin
- Mariana Bleker De Oliveira
- Chun Peng
- Laurie E. Ailles
- Geoffrey Liu
- Allan Covens
- Sergey N. Krylov
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Affiliations: Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario M3J 1P3, Canada, Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 1L7, Canada, Department of Medicine, Medical Oncology, Princess Margaret Cancer Centre, Toronto, Ontario M5G 2M9, Canada, Sunnybrook Odette Cancer Centre, Toronto, Ontario M4N 3M5, Canada
Published online on: June 14, 2021 https://doi.org/10.3892/mco.2021.2323
Article Number: 161
Copyright: © Koshkin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A primary reason for chemotherapy failure is chemoresistance, which is driven by various mechanisms. Multi‑drug resistance (MDR) is one such mechanism that is responsible for drug extrusion from the intracellular space. MDR can be intrinsic and thus, may pre‑exist the first application of chemotherapy. However, MDR may also be acquired during tumor exposure to chemotherapeutic agents. To understand whether cell clustering can influence intrinsic and acquired MDR, the present study assessed cultured monolayers (representing individual cells) and spheroids (representing clusters) formed by cisplatin‑naïve (intrinsic MDR) and cisplatin‑exposed (acquired MDR) lines of ovarian cancer A2780 cells by determining the cytometry of reaction rate constant (CRRC). MDR efflux was characterized using accurate and robust cell number vs. MDR efflux rate constant (k_MDR) histograms. Both cisplatin‑naïve and cisplatin‑exposed monolayer cells presented unimodal histograms; the histogram of cisplatin‑exposed cells was shifted towards a higher k_MDR value suggesting greater MDR activity. Spheroids of cisplatin‑naïve cells presented a bimodal histogram indicating the presence of two subpopulations with different MDR activity. In contrast, spheroids of cisplatin‑exposed cells presented a unimodal histogram qualitatively similar to that of the monolayers of cisplatin‑exposed cells but with a moderate shift towards greater MDR activity. A flow‑cytometry assessment of multidrug resistance‑associated protein 1 transporter levels in monolayers and dissociated spheroids revealed distributions similar to those of k_MDR, thus, suggesting a plausible molecular mechanism for the observed differences in MDR activity. The observed greater effect of cell clustering on intrinsic rather than in acquired MDR can help guide the development of new therapeutic strategies targeting clusters of circulating tumor cells.

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