Dados do Trabalho

Título

THE THERAPEUTIC COMBINATION OF DRUGS 5-FU AND MORPHINE REDUCES THE SURVIVAL FRACTION IN AN EXPERIMENTAL MODEL WITH PIWIL1 KNOCKOUT.

Introdução

Gastric adenocarcinoma (GA) ranks among the most aggressive tumors due to delayed diagnosis and ineffective therapeutic interventions. Dysregulation of various genes contributes to tumor malignancy, including the noteworthy alteration in the piwi protein expression (a product of the PIWIL1 gene). This altered expression can disrupt cancer-associated genes, and its heightened presence correlates with reduced survival rates, suggesting its significance in GA prognosis.

Objetivo

Given that 5-Fluorouracil (5-FU) and Morphine are integral components of the therapeutic arsenal for GA patients, this study aimed to appraise survival rates and proliferative potential following separate and combined administration of these drugs in cellular GA models influenced by PIWIL1 knockout.

Métodos

To accomplish this, AGP01 (Paraense Gastric Ascites 01) and AGP01-PIWIL1 KO (Paraense Gastric Ascites 01 with PIWIL1 gene knockout) cell lines were subjected to the Clonogenic assay with treatment durations spanning 72 hours, employing concentrations of ½ and ¼ of the IC50 for individual substances (96 and 48 μM for Morphine, 0.6 and 0.3 μM for 5-FU). Additionally, combination treatments (35 μM Morphine + 0.2 μM 5-FU, 17 μM Morphine + 0.1 μM 5-FU) were utilized, derived from the 72-hour cell viability assay via MTT. Following treatment, cells were washed and cultured for 10 days to facilitate colony formation. Colonies were captured under a stereoscopic microscope after fixation and staining and quantified using ImageJ. Survival fractions (SF) were computed as the ratio of colonies in the treated group to those in the control group (CN), then multiplied by 100. SF values underwent analysis involving the mean and standard error from 3 experiments, and comparisons were made using (2way ANOVA) and (Bonferroni) tests (p<0.05).

Resultados

Evaluation of the individual drug effects revealed that higher Morphine concentrations led to reduced SF in the AGP01 cell line expressing the PIWIL1 gene, with p<0.01. Conversely, 5-FU treatment diminished SF in both cell lines without significant intergroup variance. Intriguingly, combined treatment in the AGP01 cell line induced a considerable reduction in proliferative capacity, even at the lowest tested concentration. The difference in SF was statistically significant for both combined treatments (p<0.0001). These findings corroborate our earlier cell viability and death experiments, suggesting a synergistic effect in the cell line expressing the PIWIL1 gene.

Conclusões

Based on the furnished data, we accentuate that Morphine repositioning holds promise as a therapeutic strategy for GA, particularly in cases where PIWIL1 is upregulated; such patients experience diminished overall survival and an unfavorable prognosis.

Palavras-chave

Gastric adenocarcinoma, PIWIL1, and Cell proliferation

Financiador do resumo

Conselho Nacional de Desenvolvimento Científico e Tecnológico