Acute lymphoblastic leukemia (ALL) is the most common cancer diagnosed in pediatric age, worldwide 487,294 new cases were reported, with 305,405 deaths in both sexes in 2022. In Mexico, more than 5,000 new cases were reported. One of the main treatments for ALL is chemotherapy, which consists of a complete treatment scheme of induction, consolidation (intensification), and maintenance, to eradicate the cancer. However, after induction treatment, 10-20% of patients relapse due to chemoresistance. Therefore, it is necessary to delve deeper into the molecular mechanisms underlying chemoresistance to identify new treatment strategies to diminish or reverse it. This project focuses on the identification of proteins associated with mechanisms of chemoresistance to vincristine (VCR) and methotrexate (MTX) administered during the induction phase using proteomic (LC/MS-MS) and bioinformatic analyses. Processes related to non-response to treatment include those involving the structure and functional dynamics of the cytoskeleton, as well as nucleic acid metabolism, DNA repair, and RNA processing. TYMP and GSN were selected as differentially expressed proteins associated with these enriched processes. Validation experiments in patient samples confirm TYMP and GSN as overexpressed proteins in non-responders to vincristine and methotrexate treatment administered in the induction phase of ALL treatment. Both proteins are associated with cytoskeleton dynamics and nucleic acid metabolism, processes that are Hallmarks of chemoresistance in acute lymphoblastic leukemia.