Cell density modulates chemoresistance in breast cancer cells through differential expression of ABC transporters.

Abstract

Breast cancer patients undergoing chemotherapy encounter a significant challenge of chemoresistance because of drug efflux by ATP-binding cassette (ABC) transporters. Breast cancer cell density alters considerably throughout the early stages of primary and secondary tumor development. Although cell density in culture influences kinetics, the effects of varying cell densities on the chemoresistance of breast cancer cells remains largely unexplored. We observed chemotherapeutics-induced differential gene and protein expression of ABC transporters in luminal and basal breast cancer cells cultured at low and high seeding densities. Low-density cultures depicted a significant increase in the mRNA expression of ABC transporters-ABCG2, ABCG1, ABCC4, ABCA2, ABCA3, ABCC2, ABCC3, ABCC6, ABCC7, and ABCC9 as compared with high-density cultures. Next, cells at both low and high seeding densities when pre-treated with cyclosporine A (CsA), a pan-inhibitor of ABC transporters, resulted in increased sensitization to chemotherapeutics-doxorubicin and tamoxifen at markedly low IC50 concentrations suggesting the role of ABC transporters. Finally, markedly high doxorubicin-drug accumulation, significantly increased expression of N-cadherin, and a significant decrease in chemotherapeutics-induced in vitro tumorigenesis was observed in low-density seeded breast cancer cells when pre-treated with CsA suggesting ABC transporters inhibition-mediated increased efficacy of chemotherapeutics. These findings suggest that breast cancer cells grown at low seeding density imparts chemoresistance towards doxorubicin or tamoxifen by a differential increase in the expression of ABC transporters. Thus, a combinatorial treatment strategy including ABC transporter inhibitors and chemotherapeutics can be a way forward for overcoming the breast cancer chemoresistance.