CALN1 hypomethylation as a biomarker for high-risk bladder cancer.

Background: DNA methylation in cancer is considered a diagnostic and predictive biomarker. We investigated the usefulness of the methylation status of CALN1 as a biomarker for bladder cancer using methylation-sensitive restriction enzyme (MSRE)-quantitative polymerase chain reaction (qPCR).

Methods: Eighty-two bladder cancer fresh samples were collected via transurethral resection of bladder tumors.

Th1 polarization in the tumor microenvironment upregulates the myeloid-derived suppressor-like function of macrophages.

Here, we investigated the effect of Th1 polarization in the tumor microenvironment (TME) on tumor-associated macrophage (TAM) maturation and activation. In our immunotherapy mouse model, with a Th1-dominant TME, tumors regressed in all cases, with complete regression in 80% of the cases. Monocyte-derived dendritic cells and activated CD4 and CD8T-cells increased in the tumor-draining lymph node, and correlated with each other in the therapeutic model.

Evaluation of the efficacy of various reagents in improving microRNA extraction.

Background: MicroRNA has received considerable attention in the clinical context, and attempts are being made to use microRNA in clinical diagnosis. However, adequate quantities of microRNA required for analysis are challenging to isolate. We tested the effect of various reagents in improving microRNA extraction and compared their efficacy to that of a commercially available extraction kit (HighPure miRNA isolation kit, Roche).

Improved Target Cell Selection and Counting Method for UroVysion Fluorescence in Situ Hybridization.

Background: The UroVysion Bladder Cancer Kit requires morphological analysis of 4', 6-diamino-2-phenylindole (DAPI)-stained nuclei to identify target cells for fluorescence in situ hybridization (FISH) signals. Reproducibility and efficiency of target cell selection and counting was evaluated by combining immunofluorescence staining of cytokeratin 7 (CK7) and proliferating cell nuclear antigen (PCNA) with DAPI staining.

Methods: The reactivities to CK7, PCNA, and DAPI were compared between those for different ratios of T24 human bladder carcinoma cells and of cells from the urine of five healthy subjects.

Detection of bladder cancer by measuring CD44v6 expression in urine with real-time quantitative reverse transcription polymerase chain reaction.

Objective: To examine urinary CD44v6 total ribonucleic acid (RNA) expression in patients with bladder cancer using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and evaluate its potential as a novel marker of bladder cancer.

Methods: We used the bladder cancer cell line T24 and determined CD44v6 expression in cancer cells using in situ hybridization and immunohistochemistry. Subsequently, we obtained urine samples from 21 patients with bladder cancer and 25 patients without bladder cancer (controls).