Loss Of Adenomatous Polyposis Coli Induces Dox Resistance Through Upregulation Of Abc Transporters.

Abstract Chemoresistance is a leading cause of breast cancer related deaths. Therefore, understanding the molecular basis for chemoresistance is essential for novel therapeutic advancement improving patient outcome. The Adenomatous Polyposis Coli (APC) tumor suppressor is lost in up to 70% of sporadic breast cancer; however, little is known about how APC loss contributes to chemoresistance. Using mammary tumor cells isolated from the ApcMin/+ mouse crossed to the Polyoma middle T antigen (PyMT) transgenic model, we were the first to show that APC loss decreased doxorubicin (DOX) induced apoptosis. Therefore, we examined the mechanisms contributing to DOX resistance with APC loss to identify combination therapy options. We previously made the novel observation that APC loss in MMTV-PyMT;ApcMin/+ cells activated signal transducer and activator of transcription 3 (STAT3) thereby increasing the expression of the drug efflux pump, multidrug resistance protein 1 (MDR1). ATP-binding cassette (ABC) transporters, such as MDR1, are one of the most well-known contributors to drug resistance. These ABC transporters are capable of exporting drugs from the cell preventing apoptosis. Decreased intracellular DOX accumulation was observed in APC-deficient cells suggesting enhanced drug efflux. To investigate this decreased intracellular DOX accumulation, we measured the expression and activity of the drug export pumps, MDR1 and multidrug resistance protein 1 (MRP1). APC-deficient cells also have increased expression of MRP1. We used a calcein incorporation assay and demonstrated that APC loss increased MDR1 activity, which was restored by an MDR1 inhibitor. In addition, MDR1 inhibition sensitized the MMTV-PyMT;ApcMin/+ cells to DOX-mediated apoptosis. MRP1 inhibition through genetic manipulation will be used to assess MRP1 activity using the calcein AM assay and its contribution to DOX resistance. MDR1 and MRP1 inhibition will also be used to restore intracellular DOX accumulation. Future studies including evaluating human tumor protein lysates will for APC, MDR1, and MRP1 expression. Correlation studies will be used to determine whether loss of APC correlates with either high expression of MDR1 or MRP1. Taken together, APC loss mediates DOX resistance via exporting DOX demonstrating the potential use of combination therapy to overcome chemoresistance. Citation Format: Casey D. Stefanski, Jenifer R. Prosperi, Lauren Milac. Loss of adenomatous polyposis coli induces DOX resistance through upregulation of ABC transporters [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1404.