As a tumor involved in the urinary system, bladder cancer (BC) seriously threatens human health. Emerging as crucial biomarkers, long noncoding RNAs (lncRNAs) play an important role in the regulation of many cancers. lncRNA NNT-AS1 has been studied in a series of cancers, whereas its role and potential molecular mechanism was poorly understood in BC. Here, we found that NNT-AS1 was upregulated in BC cells. Functionally, the silencing of NNT-AS1 inhibited cell proliferation, migration, invasion, and endothelial-mesenchymal transition. Furthermore, the apoptosis of BC cells was induced upon NNT-AS1 knockdown. Later, miR-1301-3p, the downstream gene of NNT-AS1, was found at a low level in BC cells. In addition, we found that miR-1301-3p targeted to PODXL. PODXL expression downregulated in NNT-AS1-silenced cells was restored by miR-1301-3p inhibition. Importantly, NNT-AS1 was discovered to activate Wnt pathway, and the treatment of LiCl recovered the repressive role of NNT-AS1 silencing in BC cell growth. Through restoration assays, we observed that PODXL overexpressing countervailed NNT-AS1 depletionmediated suppression on BC cell growth and Wnt pathway. These data suggested that NNT-AS1 enhances BC cell growth and activates Wnt pathway by targeting miR-1301-3p/PODXL axis. K E Y W O R D Sbladder cancer, miR-1301-3p, NNT-AS1, PODXL

To select human renal cell carcinoma cell lines VMRC-RCZ After resuscitation, culture and passage, the cell concentration was adjusted to 1 × 106/ml, and the experiment was divided into a control group, a renal cancer cell + DC group (DC group), and a renal cancer cell + DC+ hTERTtransfection group (transfection Group), a total of 3 groups; the recombinant eukaryotic expression plasmid hTERT-IRES2-EGFP was constructed and transfected into mature DC; the renal cancer cell + DC group was transfected with the empty plasmid, and the renal cancer cell + DC+ hTERT transfection group was transfected with the overexpression plasmid The transfection efficiency was detected by RT-PCR method. Cultured for 12 h and 24 h respectively, the percentage of HLA-DR, CD40 and MHC-II molecules in three groups of DC immunophenotypes was determined according to flow cytometry assay, and the cytokines IL-12 and TNF-α expression level, DC apoptosis rate was detected by in situ hybridization, the rate of tumor cell proliferation was measured according to MTT method, and cell cycle was determined through flow cytometry.