Targeting doublecortin-like kinase 1 reveals a novel strategy to circumvent chemoresistance and metastasis in ovarian cancer

Targeted mRNA expression profiling was performed using nCounter® Tumor Signaling 360™ Panel (NanoString Technologies) composed of 780 genes (including internal reference genes) involved in cellular energetics, sustained proliferation, evasion of growth suppression, genomic instability, resistance to cell death, epithelial to mesenchymal transition (EMT), and metastasis. Normalization of housekeeping genes for the quantification of gene expression levels, normalization of the positive control for background correction, and data analysis were performed using the nSolverTM analysis software (NanoString Technologies). Total RNA was extracted from OVCAR-8 CPR sgCtrl and DCLK1 KO spheroids (3 resplicates/ sample). Transcripts with an RNA count < 20 for all samples were excluded because they were considered unexpressed. The mean and standard deviation (SD) were calculated for each group. Only genes with significant (p < 0.05) fold changes (< –2 or > +2) were considered differentially expressed.