The purpose of the current prospective cohort study was to determine if acculturation, measured by primary language spoken, impacts the pain response of children being treated for cancer during an experimental pain task. Sixty-seven Spanish-speaking and English-speaking children ages 6 to 18 years being treated for cancer provided ratings of pain and upset severity during the completion of the cold pressor task (CPT). One week following the CPT, participants provided their recollection of average pain and upset during the CPT. Repeated measures analysis of variance revealed Spanish-speaking children reported significantly higher pain (F1,64=5.58, P=0.02) and upset (F1,64=7.69, P=0.007) ratings during the CPT compared with English-speaking children. Also, Spanish-speaking children were over 4 times as likely to remove their hands from the water before the CPT 4-minute uninformed ceiling compared with English-speaking children (P=0.002). These findings suggest that cultural and contextual factors, including the level of acculturation, are important considerations in the assessment and management of pain in children with cancer. Future research should continue to examine the mechanisms underlying the association between acculturation and the symptom experience for children receiving treatment for cancer.