Bone invigorates metastatic seeding [RNA-seq]

We herein examine the transciptioal alteration of a single cell derived MDA-MB-231 subline,SCP21. By retrieving tumor cells inoculated in different sites, we generated multiple mammary fat pad tumor, lung metastases or bone metastasis derived SCP21 sublines. RNA-sequencing uncovered the decreased expression of EZH2 target genes in bone metastases derived cells compared other organ entrained and parental SCP21 cells , suggesting enhanced EZH2 activity. Moreover, such phenotype is reversible upon in vitro culture. As a positive control, bone derived cells were also treated with EZH2 inhibitor, EPZ011989, to confirm the decreased level of EZH2 target genes upon EZH2 inhibition. Overall design: !0E4 SCP21 cells were implanted to mammary fat pad, lung or right hindlimb of 6 weeks old female nude mice. 4 weeks later, the tissues with tumors were dissected and subjected to tumor dissociation kit. RFP+ tumor cells were sorted by flow cytometry and culture on dishes to collect enough cells for subsequent analysis. The total RNA of SCP21 cells retrieved from different sites , and cultured in vitro for different passages, or treated with EZH2 inhibitor were sequenced, in pair-end way, using Illumina NovaSeq platform.