Dados do Resumo

Título

Repositioning of Morphine in combined treatment with 5-FU as a therapeutic strategy for gastric cancer with PIWIL1 expression

Introdução

Gastric cancer (GC), the third leading cause of death globally, has adenocarcinoma as its most common type, representing 90% of cases. Dysregulation of non-coding RNAs that interact with piwi proteins (piRNAs), key regulators of gene expression, is involved in its carcinogenesis. Lower piwi expression presents better survival rates, highlighting its relevance in GC. Altered piwi expression may impact therapeutic response, emphasizing the need for new strategies, such as drug repositioning.

Objetivo

Analyze the effects of 5-FU and Morphine (MPH) on survival, proliferation, cell death patterns, and migration in GC cell models, with and without PIWIL1 gene expression, in order to investigate the influence of this gene on drug action and the potential benefits of MPH repositioning in vitro.

Métodos

For this purpose, the AGP01 (Ascite Gástrica Paraense 01), AGP01-PIWIL1 (PIWI gene knockout), and MRC-5 (pulmonary fibroblast control) human cell lines were used. These were subjected to MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) viability assays for isolated and combined drugs, with a 72-hour treatment. The selectivity index (IC) was calculated to assess the reduction in tumor cell viability compared to non-neoplastic cells. Clonogenic and cell death assays with differential staining were also performed at ½ and ¼ of the IC50 concentrations, obtained from the MTT assay, with treatment durations of up to 72 hours. The Wound Healing assay was conducted to evaluate cell migration with 24-hour treatment at the same concentrations. Statistical analyses were performed using GraphPad software, with 2-way ANOVA tests and Bonferroni post-test (p < 0.05).

Resultados

The analysis showed that the IC50 in AGP01-PIWI (MPH=288.8 µM; 5-FU=7.82 µM) was significantly higher (P=0.0012) than in AGP01 (MPH=192 µM; 5-FU=1.17 µM) and MRC-5 (MPH=143.3 µM; 5-FU=7.6 µM). Combining the drugs potentiated the IC50 in AGP01 (MPH=68.7 µM; 5-FU=0.42 µM) and AGP01-PIWI (MPH=166.9 µM; 5-FU=4.52 µM). The SI of 5-FU was higher in AGP01 (SI=6.49) compared to AGP01-PIWI (SI=0.97). MPH showed no selectivity. In combined treatment, selectivity increased in AGP01 (SI MPH=2.34; SI 5-FU=20.3). High MPH concentrations reduced the survival fraction (SF) in AGP01, while 5-FU decreased SF in both lines. Combined treatment in AGP01 reduced cell proliferation and increased apoptosis. AGP01-PIWI showed similar viable cell and apoptotics counts between combined and isolated treatments. Cell migration was significantly reduced in AGP01 and AGP01-PIWI, with 5-FU more effective at 6 hours in AGP01 and MPH, both isolated and combined, more effective in AGP01-PIWI.

Conclusões

It was observed that the combined drugs reduced viability in the PIWIL1-expressing cell line and inhibited the increased migratory capacity in both cancer cell lines. Furthermore, there was selectivity for the neoplastic cell line compared to the non-neoplastic one. Thus, it is suggested that repositioning MPH is a promising therapeutic strategy for GC with PIWIL1 expression, importat, as this pattern is associated with decreased overall survival and an unfavorable prognosis for the patient.

Financiador do resumo

National Council for Scientific and Technological Development

Palavras Chave

gastric cancer; PIWIL1; Morphine