Human bladder cancer cell line mRNA-seq

Circular RNAs (circRNAs) have been increasingly indicated to be important participants in the development and progression of various malignant tumors. Our previous studies found that hundreds of circRNAs were aberrantly expressed in bladder cancer (BC) by high-throughput sequencing and we have confirmed that the downregulated circRNAs circHIPK3, BCRC3 and circNR3C1 played inhibitory roles in BC progression. However, the role of these upregulated circRNAs remain to be clarified. In this study, we analyzed circRNA high-throughout sequencing from human tissues and focused on the upregulated circRNAs and identified a novel circular RNA, hsa_circ_0001361 (also named bladder cancer upregulated circRNA-1, BCUC-1), as a new candidate circRNA derived from FNDC3B gene. The expression levels of circRNA and miRNAs in BC tissues and cells were detected by qRT-PCR. Transwell migration, Matrigel invasion, CCK-8, EdU, cell cycle and in vivo tumor metastasis assays were preformed to evaluate the effects of circ0001361 on BC cells. Transcriptome analysis was performed to find the potential genes that could be regulated by BCUC-1. Luciferase reporter, RNA pull-down and fluorescence in situ hybridization (FISH) assays were applied to verify the interaction between BCUC-1 and miR-491-5p. Overall design: In order to screen downstream genes regulated by BCUC-1 (hsa_circ_0001361), transcriptome analysis was performed in T24T and UMUC3 cells transfected with BCUC-1(has_circ_0001361) and compaired vector.