Cancer Therapeutics Insights Tumor-Initiating Cells and FZD8 Play a Major Role in Drug Resistance in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) studies have shown that neoadjuvant chemotherapy before surgery was effective in the minority of women, whereas the majority who had residual tumor had a relatively poor outcome. To identify the mechanism by which residual cancer cells survive chemotherapy, we initially conducted gene expression profiling using the CRL2335 TNBC cell line derived from a squamous breast carcinoma before and after treatment with cisplatin plus TRAIL. We found a significant increase in the expression of FZD8, one of Wnt receptors, and its downstream targets LEF1 and TCF in residual CRL2335 tumor cells after treatment with cisplatin plus TRAIL. Increased FZD8 levels were further confirmed in other TNBC cell lines. Inhibition of FZD8 by siRNA inCRL2335 cells in the presence of cisplatin plus TRAIL reduced b-catenin and survivin levels and increased apoptosis compared with scrambled siRNA–treated cells. In vivo data show that cisplatin plus TRAIL treatment significantly reduces tumor volume in NOD/SCID mice. However, we found that cisplatin plus TRAIL treatment predominantly eliminated non–tumor-initiating cells, as shown by whole-body fluorescent imaging of mice injected with mammosphere-forming CRL2335 cells stably transfectedwithDsRed. This led toTIC enrichment in residual tumors, as confirmedby immunostaining for TIC markers. Moreover, an increase in FZD8 expression was observed in residual tumors treated with cisplatin andTRAIL. Taken together, ourfindings suggest that FZD8-mediatedWnt signalingmayplay amajor role inmediating resistance to chemotherapy,making it apotential target to enhance chemotherapeutic efficacy in patients with TNBCs. Mol Cancer Ther; 12(4); 491–8. 2013 AACR.