Although targeting programmed death 1/programmed death ligand 1 (PD-1/PD-L1) has achieved durable responses and disease remission in patients with certain cancers, relatively low response rates and emerging resistance limit its clinical application. Hence, a more thorough understanding of regulatory mechanisms of the PD-1/PD-L1 axis is vital for developing combined therapeutic strategies to overcome hurdles of PD-1/PD-L1 blockade. Increasing evidence has demonstrated that PD-L1 can be secreted into the extracellular space or translocated into the nucleus, which also plays a critical role in regulating cancer immune evasion, tumorigenesis, and immunotherapy. In this review, we summarize these emerging roles of extracellular and nuclear PD-L1 and discuss future research directions and potential opportunities in translational medicine.