Recent studies suggest that metformin, a widely used antidiabetic agent, may reduce cancer risk and improve prognosis of certain malignancies. However, the mechanisms for the anti-cancer effects of metformin remain uncertain. In this study, we investigated the effects of metformin on human bladder cancer cells and the underlying mechanisms. Metformin significantly inhibited the proliferation and colony formation of 5637 and T24 cells in vitro; specifically, metformin induced an apparent cell cycle arrest in G0/G1 phases, accompanied by a strong decrease of cyclin D1, cyclin-dependent kinase 4 (CDK4), E2F1 and an increase of p21^waf-1. Further experiments revealed that metformin activated AMP-activated protein kinase (AMPK) and suppressed mammalian target of rapamycin (mTOR), the central regulator of protein synthesis and cell growth. Moreover, daily treatment of metformin led to a substantial inhibition of tumor growth in a xenograft model with concomitant decrease in the expression of proliferating cell nuclear antigen (PCNA), cyclin D1 and p-mTOR. The in vitro and in vivo results demonstrate that metformin efficiently suppresses the proliferation of bladder cancer cells and suggest that metformin may be a potential therapeutic agent for the treatment of bladder cancer.

The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), either alone or in combination with other anti-cancer agents, has been considered as a new strategy for anti-cancer therapy. In this study, we demonstrated that evodiamine, a quinolone alkaloid isolated from the fruit of Evodia fructus, induced apoptosis and enhanced TRAIL-induced apoptosis in human bladder cancer cells. To elucidate the underlying mechanism, we found that evodiamine significantly reduced the protein levels of Mcl-1 in 253J and T24 bladder cancer cells, and overexpression of this molecule attenuated the apoptosis induced by evodiamine alone, or in combination with TRAIL. Further experiments revealed that evodiamine did not affect the mRNA level, proteasomal degradation and protein stability of Mcl-1. On the other hand, evodiamine inhibited the mTOR/S6K1 pathway, which usually regulates protein translation; moreover, knockdown of S6K1 with small interfering RNA (siRNA) effectively reduced Mcl-1 levels, indicating evodiamine downregulates c-FLIP through inhibition of mTOR/S6K1 pathway. Taken together, our results indicate that evodiamine induces apoptosis and enhances TRAIL-induced apoptosis possibly through mTOR/S6K1-mediated downregulation of Mcl-1; furthermore, these findings provide a rationale for the combined application of evodiamine with TRAIL in the treatment of bladder cancer.

Background: The development of benign prostatic hyperplasia (BPH) is closely related to hypoxia in the prostatic stroma, and the hypoxia-inducible factor-1α/vascular endothelial growth factor (HIF-1α/VEGF) pathway has been shown to significantly activate in response to hypoxia. The underlying mechanism for activation of this pathway in the pathogenesis of BPH remains unclear. Materials and methods: We constructed HIF-1α overexpression and knockdown BPH stromal (WPMY-1) and epithelial (BPH-1) cell lines, which were cultured under different oxygen conditions (hypoxia, normoxia, and hypoxia + HIF-1α inhibitor). Quantitative real-time polymerase chain reaction (qPCR) and Western blotting were applied to detect the expression of the HIF-1α/VEGF pathway. Cell proliferation and apoptosis were analyzed by Cell Counting Kit-8 and flow cytometry. We used the miRWalk 2.0 database and Western blotting to predict the potential miRNA that selectively targets the HIF-1α/VEGF pathway, and verified the prediction by qPCR and dual-luciferase assays. Results: In a BPH stromal cell line (WPMY-1), the expression of VEGF was in accordance with HIF-1α levels, elevated in the overexpression cells and decreased in the knockdown cells. Hypoxia-induced HIF-1α overexpression, which could be reversed by a HIF-1α inhibitor. Moreover, the HIF-1α inhibitor significantly depressed cellular proliferation and promoted apoptosis in hypoxic conditions, assessed by Cell Counting Kit-8 and flow cytometry. However, in the BPH epithelial cell line (BPH-1), the expression level of HIF-1α did not influence the expression of VEGF. Finally, a potential miRNA, miR-17-5p, regulating the HIF-1α/VEGF pathway was predicted from the miRWalk 2.0 database and Western blotting, and verified by qPCR and dual-luciferase assay. Conclusions: In hypoxia, activation of the HIF-1α/VEGF pathway plays a crucial role in regulating cell proliferation in a BPH stromal cell line. Regulation by miR-17-5p may be the potential mechanism for the activation of this pathway. Regulation of this pathway may be involved in the pathogenesis of BPH.

To study the effect and predict the value of neoadjuvant treatment regimen for muscle invasive bladder cancer (MIBC) by construction of a columnar graphical model of patients by neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), and systemic immune-inflammatory index (SII) indexes. 265 patients with MIBC included from May 2022 to May 2024 were retrospectively selected to receive neoadjuvant treatment regimen respectively with treatment effect assessed, among which those achieving complete response (CR), partial response (PR), or stable disease (SD) were included in responders group and those with progressive disease (PD) in non-responders group. Clinical data of both groups were compared, related factors affecting the poor outcome after neoadjuvant therapy for MIBC were analyzed by Logistic regression, ensued with analysis of predictive value of poor prognosis by construction of a columnar graph model based on the NLR, dNLR and SII indexes. A total of 265 patients with MIBC were included in this paper with a disease control rate (DCR) of 84.53% (224/265) after treatment with neoadjuvant regimen, among which 224 cases with controlled disease were involved in responders group and the remaining 41 cases with PD in non-responders group. Significant differences were observed between the two groups in terms of the degree of differentiation, tumor stage, NLR, dNLR and SII index levels (P < 0.05). After the diagnosis of covariance, the VIF values of the degree of differentiation and tumor stage were 5.535 and 5.582 respectively with a tolerance of 0.181 and 0.179, indicating that there existed a covariance problem (VIF value > 5) and could be moved out of the model followed by secondary analysis. Variables with P < 0.05 in the univariate factors were involved in the multivariate Logistic regression model with results showing that NLR, dNLR, and SII were all influential factors for the poor outcome of neoadjuvant regimens after treatment of MIBC (P < 0.05). Next, the column line graph, calibration curve and ROC curve graph were constructed. It was found that the AUC of the column line graph model in predicting poor outcome after neoadjuvant regimen for MIBC registered 0.995 (95% CI: 0.99-1.00), which was valuable in predicting poor outcome after neoadjuvant regimen for MIBC. CYFRA21-1, NMP22, and BTA were significantly higher in the poor response group than in the response group (P < 0.05), and CYFRA21-1, NMP22, and BTA showed a positive correlation with NLR, dNLR, and SII in both groups, respectively (P < 0.05). The neoadjuvant treatment program in patients with MIBC performed better, but some patients might still have a poor outcome with higher levels of NLR, dNLR and SII compared to those with a good outcome. In addition, the value of the combination of the three indicators in the prediction of the neoadjuvant treatment program displayed better performance, which was able to provide reference value for clinical decision-making.

Background Clear cell renal cell carcinoma (ccRCC) poses significant challenges due to its asymptomatic nature and poor prognosis at advanced stages. Identifying novel biomarkers is essential for enhancing prognostic accuracy and therapeutic strategies. This study explores the CWH43 gene, utilizing multi-omics data to determine its role in ccRCC. Methods Genomic, transcriptomic, and methylation data from TCGA-KIRC and GEO databases were analyzed to evaluate CWH43 expression and clinical impact. Bioinformatics tools assessed correlations with patient outcomes and pathway involvement. Results CWH43 expression was significantly reduced in ccRCC tissues and correlated with advanced disease stages and poor patient survival. Enrichment analyses revealed CWH43's involvement in critical cancer pathways, such as autophagy and immune response modulation, suggesting its significant role in ccRCC pathophysiology. Lower CWH43 levels were associated with increased tumor progression and immune evasion, impacting the tumor microenvironment. Conclusion This study highlights the utility of multi-omics data in identifying CWH43 as a novel prognostic biomarker for ccRCC. Integrating CWH43 into clinical practice could refine prognostic assessments and guide personalized therapy strategies, aligning with advancements in modern oncology. Further research is warranted to explore CWH43's mechanisms and therapeutic potential.