This study explores the long-term adaptation mechanisms of lymphoma cells subjected to hypoxic conditions, with an emphasis on the HBL2 and Ramos cell lines under normoxia and 1% O2 environments. The research methodology encompassed lysing cells using a buffer containing sodium deoxycholate and TEAB, followed by protein quantification via a BCA assay. Subsequent steps involved protein digestion with trypsin, labelling with TMTpro™ 16plex Label Reagent Set for quantitative analysis, peptide fractionation, and LC-MS/MS analysis. The analytical process was supported by Proteome Discoverer 2.4 for protein identification and quantification. Experimental groups were categorized into "HBL2 normoxia," "HBL2 1% O2 adaptation," "Ramos normoxia," and "Ramos 1% O2 adaptation," conducted in triplicates to ensure reliability. This meticulous approach facilitated the delineation of unique proteomic landscapes indicative of hypoxia adaptation, unveiling cellular strategies employed by lymphoma cells to navigate low oxygen conditions. The findings advance our comprehension of how hypoxia influences cancer progression and potentially opens new avenues for targeting hypoxic niches within tumors.