Modified Nucleosides as Potential Biomarkers of Prostate Cancer: Targeted Metabolomics of In Vitro Cell Samples by MEKC‐UV

ABSTRACT Prostate cancer is the second most common cancer among men globally, with over 1.4 million new cases and nearly 400000 deaths reported in 2022. Despite the availability of diagnostic tools such as the Prostate Specific Antigen (PSA) test, its low sensitivity reinforces the need for the exploration of more reliable biomarkers. In this context, metabolomics offers a promising approach for identifying sensitive biomarkers to improve cancer diagnosis and treatment. Therefore, this study aimed to conduct a targeted metabolomic analysis of the extracellular environment of In Vitro non‐tumoral and cancer prostate cells to compare the levels of eight nucleosides using micellar electrokinetic capillary chromatography with UV detection (MEKC‐UV). The method was adapted from a previously optimized protocol for blood serum, with minor adjustments to meet the Brazilian National Health Surveillance Agency (ANVISA) standards. Nucleosides were extracted via solid‐phase extraction (SPE), and cell cultures were maintained under controlled conditions at 37°C with 5% CO 2 until reaching 80% confluence. The optimized MEKC‐UV method demonstrated precision and accuracy, although the Youden test indicated some lack of robustness. Statistical analysis using a two‐tailed t ‐test revealed significantly higher adenosine levels in non‐tumoral cells, whereas uridine and 5‐methyluridine concentrations were elevated in cancer cells. Inosine was detected exclusively in the non‐tumoral cell line. Nevertheless, the method's innovative and cost‐effective nature underscores its potential as a tool for cancer biomarker identification, with distinct nucleoside patterns in cancer cells offering valuable insights for disease recognition.