Análise do perfil ultraestrutural e molecular das linhagens MNP-01 e AGP-01: identificação de biomarcadores associados ao fenótipo maligno no câncer gástrico

Abstract

Gastric cancer (GC) is a global public health challenge, ranking fifth in both incidence and mortality among malignant neoplasms. Its development is multifactorial, involving genetic, epigenetic, biochemical, and morphological alterations. Considering the scarcity of studies in Brazilian populations, this study aimed to compare the ultrastructural and molecular profiles of the gastric cell lines MNP-01 (non-tumoral) and AGP-01 (tumoral), in order to prospect protein biomarkers associated with the malignant phenotype. Ultrastructural and nanomechanical parameters were evaluated by AFM, while protein and other biomolecule profiles were analyzed by shotgun proteomics (UHPLC-MS/MS) and Raman spectroscopy, respectively. AFM maps revealed differences in the cell surface: the tumor cell line AGP-01 exhibited lower roughness, tumor volume, and surface area, as well as greater deformability and reduced Young’s modulus and adhesion values. In turn, Raman spectroscopy revealed differences in the vibrational profile as well as a distinct molecular composition, including nucleic acids, lipids, and proteins. Proteomics identified 2,209 proteins, of which 433 were exclusive to the MNP-01 lineage, 328 to the AGP-01 lineage, and 724 shared between both. From this total, proteins involved in protein synthesis and translation, oxidative stress and metabolism, cell cycle regulation, cell adhesion, drug resistance, and cytoskeletal reorganization were highlighted. Protein interaction and functional analyses indicated alterations in oncogenic pathways, especially AKT/mTOR, which were confirmed by immunofluorescence. The tumoral lineage AGP-01 showed a significant increase in p-AKT (1.8× vs. MNP-01; p = 0.0289) and p-mTOR (3× vs. MNP-01; p = 0.004), indicating hyperactivation of this pathway as a feature of the malignant phenotype. Altogether, the results demonstrate that the AGP-01 tumor cell line is associated with the integration of ultrastructural alterations, proteomic reprogramming, and activation of pro-survival pathways, broadening the understanding of tumor biology and the biomechanical and molecular bases of metastatic GC, providing support for the identification and investigation of potential therapeutic targets, as well as for future studies with functional validation and translational application.