<p>Lenalidomide (LEN) is a mainstay for treating multiple myeloma (MM), but its efficacy is often limited by resistance. We investigated the interaction between aldo–keto reductase family 1 member C3 (AKR1C3) and Yin Yang 1 (YY1) and their roles in LEN resistance. We induced LEN-resistant MM cell lines (H929R and U266R). Loss- or gain-of-function assays of AKR1C3 and YY1 were used to analyse the half maximal inhibitory concentration (IC₅₀) values, cell senescence, DNA damage, and glycolytic activity under LEN treatment. Chromatin immunoprecipitation was used to determine the interaction between YY1 and AKR1C3. As results, AKR1C3 and YY1 were upregulated in H929R and U266R cells. AKR1C3 silencing decreased the LEN’s IC₅₀, slowed cell growth, enhanced senescence and DNA damage, reduced metabolic reprogramming. YY1 activated the transcription of AKR1C3 by binding to its promoter region. Similarly, silencing YY1 enhanced LEN sensitivity, suppressed glycolysis, and was counteracted by AKR1C3 overexpression. Mechanistically, YY1-AKR1C3 activated the Hedgehog pathway; fluticasone reversed the effects of AKR1C3 silencing on LEN resistance and glycolysis in H929R and U266R cells. Overall, YY1 activates AKR1C3 transcription and the Hedgehog pathway to increase LEN resistance and glycolytic activity in MM cells.</p>