Impact of HGFL-Ron signaling on breast cancer stem cell transcriptomic profiles.

Introduction: HGFL-Ron signaling is augmented in human breast cancer and is associated with poor overall prognosis. Here, we investigate the role of HGFL-Ron signaling in murine breast cancer stem cells (BCSC) through characterization of BCSC transcriptomes through RNA-sequencing, focusing on the impact of Ron knockdown through a short hairpin construct. Methods:R7 breas cancer cell lines were drived from mammary tumors in transgenic MMTV_Ron mice. They were sorted based on expression of cell surface markers indicative of lineage negative, CD29hi and CD24+ cells. Bulk R7, sorted cells, and sorted cells treated with shRon were submitted for transcriptomic characterization on the Illumina HiSeq 2500. High quality reads were aligned to the mm9 genome and quantified to generate RPKM. Results: Approximately 30 million reads were aligned to the mouse genome in each sample which corresponded to over 36000 transcripts. Of these, ~16000 were included in analysis. Conclusions: Differential expression analysis indicated that depletion of Ron markely reduces mammosphere formation and self-renewal, and highlighted by the decrease in B-catenin and NFKB pathways. Overall design: Transcriptome profiles of bulk and sorted R7 BCSCs with Ron knockdown through RNA-sequencing.

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Ruiz-Torres SJ, Benight NM, Karns RA, Lower EE, Guan JL, Waltz SE