Objective. To verify and evaluate the value of CD40 as a noninvasive biomarker of cisplatin-resistant bladder cancer, we studied the expression of CD40 and the correlation between nucleophosmin (NPM1) and CD40 in cisplatin-resistant bladder cancer. Methods. Three cisplatin-resistant bladder cancer cell lines (T24/0.8DDP, BIU87/0.3DDP, and PUMC-91/0.6DDP) were studied, and lentivirus was used to silence NPM1 expression. The expression of CD40 and NPM1 in three NPM1 silencing bladder cancer cell lines were detected by fluorescence microscopy and Western Blot. The effects and proteomic bioinformatics of NPM1 gene knockout on cisplatin-resistant bladder cancer cells were analyzed by liquid chromatography-mass spectrometry (LC-MS) and gene ontology analysis (GO analysis). Results. The NPM1 gene was successfully silenced in three drug-resistant bladder cancer cell lines by lentivirus infection. The knockdown efficiency was 70%. After NPM1 gene knockout, 492 differential proteins were detected by LC-MS, whose fold change was more than 1.5 p<0.05. A total of 57022 peptides, 54347 unique peptides, and 6686 protein groups were identified in all proteins of the tested cells (FDR < 0.01). Hierarchical clustering and principal component analysis showed that 264 functional proteins were downregulated and 228 functional proteins were upregulated in the gene silencing group compared with those of the negative controls. By GO analysis, the proteins affected by NPM1 cover a large number of proteins with biological functions, which may play an important role in the development of tumor in 492 differential proteins. The CD40 was the most significantly downregulated protein after NPM1 silencing, with a difference of 2.6-fold change in abundance. Conclusions. There is a positive correlation between CD40 protein and NPM1 protein in drug-resistant bladder cancer. Because NPM1 can reflect the characteristics of bladder cancer, CD40 may be a more sensitive marker for monitoring the prognosis of bladder cancer.

Background: Ferritin plays a key role in the development of prostate cancer (PCa). Our earlier studies showed that the knockdown of ferritin heavy chain (FTH) suppressed the migration and invasion of the prostate cancer cell line (PC3). However, the mechanisms behind FTH in the cell migration regulation of PCa have not been thoroughly investigated.Methods: Isobaric tags for relative and absolute quantitation (iTRAQ) proteomics was used to analyze the protein expression in PC3 cells with FTH knockdown by small interfering RNAs and negative control cells.We subsequently ranked the differentially expressed proteins according to the change in expression. We further performed Gene Ontology (GO) analysis for the changing-expression protein. Finally, Western blot analysis was performed to determine the expression of the target protein.Results: Compared with the negative group, 420 proteins were downregulated, including proteins S100A4, S100P, and S100A2, while the expression of 442 protein was elevated in FTH-silencing PC3 cells (P<0.05, fold change >1.2). The mass spectrometry results showing decreased expression of protein S100A4, S100P, and S100A2 in the cells were further validated by Western blot (P<0.05). Levels of protein S100A4, S100A2, and S100P were reduced in FTH-silencing PC3 cells (P<0.05, fold change >1.6). Conclusions:The downregulation of FTH expression reduced the level of protein S100A4, S100A2, and S100P, which all play a key role in the migration and invasion of tumor cells. Therefore, it is reasonable to assume that there are correlations between the expression of the S100A4, S100A2, and S100P genes with FTH. Based on this research, FTH may be a new biomarker for the diagnosis of PCa.

Background: NPM1 can provide abundant information of bladder cancer changes, but the effect of NPM1 differential expression on tumor and tumor related molecular mechanism has not been elucidated.Methods: NPM1 silencing cell line was established by lentivirus. The tumorigenic ability was judged by wound-healing assay, transwell invasion assay and nude mice tumorigenicity assay. The proteome of NPM1 deficient bladder cancer cell line was analyzed by Liquid Chromatography Mass Spectrometry (LC-MS). The results of mass spectrometry are comprehensively analyzed by bioinformatics analysis for tumor related molecules. The signal pathways involved in tumor related molecules will be verified by KEGG and UniProt databases.Results: Compared with the corresponding negative control group, NPM1 silencing cell line T24/DDP Lv-NPM1 showed strong migration ability and high invasive ability. There was no significant difference in migration ability and the invasive cells proportion between NPM1 overexpressing cell line and related negative control group. The tumorigenesis in nude mice also showed that NPM1 silencing tumor had larger tumor volume. Among all differential proteins analyzed by mass spectrometry, 20 proteins with signal transduction activity showed the most significant difference (Fold change > 1.5). 6 of them were associated with NF-κB signaling pathway, which may play an important role in the development of tumor.Conclusions: The loss of NPM1 may indicate the poor outcome of bladder cancer. Abnormal expression of NF-κB signaling pathway is an important factor in the progression of bladder cancer. Monitoring NPM1 expression can effectively adjust the treatment strategy of bladder cancer.