BAP1-Related ceRNA (NEAT1/miR-10a-5p/SERPINE1) Promotes Proliferation and Migration of Kidney Cancer Cells

Abstract

BackgroundBAP1 is an important tumor suppressor involved in various biological processes and is commonly lost or inactivated in clear-cell renal cell carcinoma (ccRCC). However, the role of the BAP1-deficient tumor competing endogenous RNA (ceRNA) network involved in ccRCC remains unclear. Thus, this study aims to investigate the prognostic BAP1-related ceRNA in ccRCC.MethodsRaw data was obtained from the TCGA and the differentially expressed genes were screened to establish a BAP1-related ceRNA network. Subsequently, the role of the ceRNA axis was validated using phenotypic experiments. Dual-luciferase reporter assays and fluorescence in situ hybridization (FISH) assays were used to confirm the ceRNA network.ResultsNuclear enriched abundant transcript 1 (NEAT1) expression was significantly increased in kidney cancer cell lines. NEAT1 knockdown significantly inhibited cell proliferation and migration, which could be reversed by miR-10a-5p inhibitor. Dual-luciferase reporter assay confirmed miR-10a-5p as a common target of NEAT1 and Serine protease inhibitor family E member 1 (SERPINE1). FISH assays revealed the co-localization of NEAT1 and miR-10a-5p in the cytoplasm. Additionally, the methylation level of SERPINE1 in ccRCC was significantly lower than that in normal tissues. Furthermore, SERPINE1 expression was positively correlated with multiple immune cell infiltration levels.ConclusionsIn BAP1-deficient ccRCC, NEAT1 competitively binds to miR-10a-5p, indirectly upregulating SERPINE1 expression to promote kidney cancer cell proliferation. Furthermore, NEAT1/miR-10a-5p/SERPINE1 were found to be independent prognostic factors of ccRCC.

Yin Yang 1 promotes the neuroendocrine differentiation of prostate cancer cells via the non‐canonical WNT pathway (FYN/STAT3)

“…Colony formation was assessed by seeding cells into 6‐well plates (500 cells/well) in triplicate and culturing for 10 days, after which the colonies were fixed, stained, and counted. For the EdU assay kit (RiboBio), cells were seeded into 24‐well plates (3 × 10 4 cells/well) in triplicate, cultured for 24 h, then incubated with 50 μM EdU for 2 h. Subsequent staining and visualization were performed 76 . The EdU assay involves the addition of a penetrant to cell climbing slides that have been prepared and subsequently incubated in a rocker device for 10 min.…”

“…For the EdU assay kit (RiboBio), cells were seeded into 24‐well plates (3 × 10 4 cells/well) in triplicate, cultured for 24 h, then incubated with 50 μM EdU for 2 h. Subsequent staining and visualization were performed. 76 The EdU assay involves the addition of a penetrant to cell climbing slides that have been prepared and subsequently incubated in a rocker device for 10 min. Subsequently, the EdU staining reaction solution was added and incubated at room temperature for 30 min in the dark.…”

Coordinated reprogramming of renal cancer transcriptome, metabolome and secretome associates with immune tumor infiltration

“…SERPINE1 is a well-known component of human secretome involved in senescence, fibrosis, and cancerous progression [ 51 – 53 ], released by cancer cells [ 52 , 54 – 56 ] and TME components including adipocytes, osteoblasts, CAFs, and MSCs [ 57 – 61 ]. High SERPINE1 expression correlates with the immune infiltration in ccRCC tumors [ 62 ] and poor prognosis for ccRCC patients [ 63 ]. SERPINE1 acts in an autocrine manner, contributing to the motility of ccRCC cells [ 64 ].…”

Advances in targeting of miR‑10‑associated lncRNAs/circRNAs for the management of cancer (Review)

“…The area under the curve (AUC) of the receiver operating characteristic curve analysis indicated that SERPINE1 (AUC=0.789) and miR-10a-5p (AUC=0.892) had good prognostic performance. Furthermore, SERPINE1 expression was positively associated with the levels of immune infiltration of CD4 + T cells, CD8 + T cells, macrophages, dendritic cells and neutrophils ( 82 ). In another study, a ceRNA regulatory network consisting of the lncRNA MMP25-AS1/hsa-miR-10a-5p/SERPINE1 axis was identified.…”