Translatome and transcriptome sequencing data of 16-day kinetic response in colorectal cancer to 5-FU chemotherapy

Resistance to 5-fluorouracil (5-FU) remains a major challenge in colorectal cancer therapy, often resulting in tumor relapse fueled by drug-tolerant persister (DTP) cells. Unexpectedly, instead of suppressing translation under chemotherapeutic stress, 5-FU by integrating ribosomal RNA of persister cells drives a transient surge in protein synthesis, revealing a distinct adaptive state. This translational reprogramming drives a highly plastic subpopulation that supports survival under drug pressure. Integrated translatome profiling uncovered selective translation of mRNAs linked to survival pathways including epigenetic and metabolic changes, apoptosis resistance, cell cycle arrest and a senescence-like phenotype. Together, our findings uncover active translation as an unrecognized hallmark of the DTP state. Overall design: Comparison of cytoplasmic transcriptome and polysome profiling fractions (more than 3 linked ribosomes) of HCT116 colorectal cancer cells treated with 5-FU. RNA samples were collected at 6 time points (D0, D1, D2, D7, D14 and D16) after treatment, 3 replicates were used.