Targeting the Kynurenine Pathway in an In vitro Model of Pancreatic Ductal Adenocarcinoma
Project no. 23636 · Status: Kapanmış · 2022
Summary
<p>In our study, based on the analysis we conducted in the TCGA PAAD cohort, we found that the mRNA expression of KYNU, which encodes the kynureninase enzyme, was significantly higher in pancreatic cancer tumor tissues compared to normal pancreatic tissues (ntumor=179, nnormal=171, p<0.01). We also demonstrated that increased expression was significantly associated with decreased overall survival in patients (Logrank p=0.0029; HR=2.2) and was associated with TNM staging (Pr(>F)=0.0052). Subsequently, in an in vitro model of pancreatic cancer, we evaluated the effects of pharmacological inhibitors, Ro 61-8048 and 3-Hydroxyhippuric acid (3-HHA), on cell viability, cell migration, and intracellular NAD+/NADH ratio using crystal violet cell viability assay, wound healing assay, and spectrophotometry, respectively. The study results indicate that both Ro 61-8048 and 3-HHA treatments have the capacity to inhibit cell viability in pancreatic cancer moderately. Regarding migration inhibition, Ro 61-8048 treatment showed a higher inhibition rate than the 3-HHA treatment. According to the analysis of NAD+ and NADH, no statistically significant change was observed in cells treated with either Ro 61-8048 or 3-HHA alone compared to the DMSO control group. Our results indicated that Ro 61-8048 and 3-HHA single treatments have a modest capacity to inhibit cell viability in pancreatic cancer. Ro 61-8048 treatment was found to provide a 52.40% migration inhibition compared to the DMSO control group (p=0.0003), while the single 3-HHA treatment inhibited the migration by 26.53% compared to the DMSO control group (p=0.0096). There was no statistically significant change in NAD+/NADH ratio and levels in cells treated with Ro 61-8048 and 3-HHA compared to DMSO control groups. The data obtained as a result of this study revealed that KYNU mRNA expression has the potential to be both a diagnostic and prognostic biomarker in pancreatic cancer and provided evidence that Ro 61-8048 and 3-HHA treatments may suppress migration in pancreatic cancer cells without affecting intracellular NAD+ levels. Validation of the obtained results in in vivo studies may pave the way for developing new strategies for suppressing metastasis, which is one of the significant factors limiting the success of pancreatic cancer treatment.</p>