Background <p>Healthcare professionals’ perceptions of palliative care (PC) vary and are shaped by culture, educational experiences, and access to resources.</p> Objective <p>This study aimed to identify self-reported perceptions and knowledge gaps regarding PC among healthcare professionals in a Brazilian university hospital, aligned with the National Humanization Policy of the Unified Health System (SUS).</p> Methods <p>An observational quantitative study used a self-administered structured questionnaire administered to physicians, nurses, social workers, physical therapists, nutritionists, psychologists, nursing technicians, and other professionals.</p> Results <p>About one quarter of participants were unaware of the hospital’s PC service, with significant differences across professions (<i>p</i> &lt; 0.001). Key barriers included prognostic uncertainty, limited conceptual understanding of PC, and concerns about patient and family acceptance, consistent with international reports. Participants agreed that broader professional involvement in end-of-life discussions is needed and that PC services were underutilized (<i>p</i> = 0.003), mainly because of limited access to or adherence to information. </p> Conclusions <p>Differences in perceptions regarding PC referral and communication were observed according to participants’ reported educational experiences. Confusion also persisted regarding the appropriate timing for initiating PC. Targeted awareness initiatives, interprofessional education, and improved institutional communication are needed to strengthen implementation and timely access to PC in routine clinical practice.</p>