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. !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.