A growing number of dysregulated long non-coding (lnc)RNAs have been verified to serve an essential role in human prostate cancer. However, the underlying mechanisms of lncRNA MNX1 Antisense RNA 1 (MNX1-AS1) in prostate cancer has not been explored. Therefore, the present study aimed to explore the function of MNX1-AS1 in prostate cancer tumorigenesis and investigate the in-depth mechanism. The expression of MNX1-AS1, microRNA (miR)-2113 and murine double min 2 (MDM2) in prostate cancer tissues and corresponding normal tissues were assessed by reverse transcription-quantitative PCR. The protein expression levels of MDM2 were detected by western blotting. LNCaP and PC-3 cells were transfected with short hairpin (sh)-MNX1-AS1, miR-2113 mimics, miR-2113 inhibitor and pCDH-MDM2 vector using Lipofectamine ® 3000. Cell proliferation, migration and invasion abilities were assessed by CCK-8 assay, colony formation and Transwell assay, respectively. Dual luciferase reporter assay was carried out to confirm the putative targets of MNX1-AS1 and miR-2113. Tumor formation experiment in nude mice was applied to evaluate the tumor growth effect of MNX1-AS1 in vivo . The expression of MNX1-AS1 was significantly upregulated in the prostate cancer tissues and cell lines. MNX1-AS1 knockdown suppressed the abilities of cell viability and migration and invasion in vitro and inhibited tumor growth in vivo . Additionally, luciferase reporter assay revealed that MNX1-AS1 could target miR-2113 and negatively interacted with miR-2113 in prostate cancer cells. miR-2113 directly targeted to MDM2 and negatively modulated the expression of MDM2. Rescue assays suggested that the viability, migration and invasion of impaired cells triggered by transfection with sh-MNX1-AS1 alone could be recovered by co-transfection with sh-MNX1-AS1 + miR-2113 inhibitor or sh-MNX1-AS1 + pCDH- MDM2 vector. The present study demonstrated that MNX1-AS1 promoted prostate cancer progression through regulating miR-2113/ MDM2 axis.

Biochemical recurrence (BCR) is a cause of concern in advanced prostate cancer (PCa). Thus, novel diagnostic biomarkers are required to improve clinical care. However, research on PCa immunotherapy is also scarce. Hence, the present study aimed to explore promising BCR-related diagnostic biomarkers, and their expression pattern, prognostic value, immune response effects, biological functions, and possible molecular mechanisms were evaluated. GEO datasets (GSE46602, GSE70768, and GSE116918) were downloaded and merged as the training cohort, and differential expression analysis was performed. Lasso regression and SVM-RFE algorithm, as well as PPI analysis and MCODE algorithm, were then applied to filter BCR-related biomarker genes. The CIBERSORT and estimation of stromal and immune cells in malignant tumor tissues using expression data (ESTIMATE) methods were used to calculate the fractions of tumorinfiltrating immune cells. GO/DO enrichment analyses were used to identify the biological functions. The expression of latent transforming growth factor β-binding protein 2 (LTBP2) was determined by RT-qPCR and western blotting. The role of LTBP2 in PCa was determined by CCK-8, Transwell, and the potential mechanism was investigated by KEGG and GSEA and confirmed by western blotting. In total, 44 BCR-related differentially expressed genes (DEGs) in the training cohort were screened. LTBP2 was found to be a diagnostic biomarker of BCR in PCa and was associated with CD4 + T-cell infiltration and response to anti-PD-1/PD-L1 immunotherapy. Subsequently, using the ESTIMATE algorithm, it was identified that LTBP2 was associated with the tumor microenvironment and could be a predictor of the clinical benefit of immune checkpoint blockade. Finally, the expression and biological function of LTBP2 were evaluated via cellular experiments. The results showed that LTBP2 was downregulated in PCa cells and inhibited PCa proliferation and metastasis via the PI3K/AKT signaling pathway in vitro. In conclusion, LTBP2 was a promising diagnostic biomarker of BCR of PCa and had an important role in CD4 + T-cell recruitment. Moreover, it was associated with immunotherapy in patients with PCa who developed BCR, and it inhibited PCa proliferation and metastasis via the PI3K/AKT signaling pathway in vitro.