Peroxiredoxin IV (Prx IV), a member of the peroxiredoxin family, has been shown to be involved in cell protection against radiation. Peroxiredoxins are also overexpressed and involved in the progression of several tumours. Calpains have been shown to be over-activated in alveolar rhabdomyosarcoma (ARMS). The present study focused on the possible cross-regulations between Prx IV and calpains in ARMS cells. Prx IV abundance was quantified by Western blot analysis in ARMS cells and compared with non-malignant LHCN-M2 cells. Its abundance is quantified in ARMS cells treated or untreated with calpain inhibitors moreover its mRNA expression is also quantified by real-time RT-PCR in these cells. The study showed that Prx IV is overexpressed by five times in ARMS cells when compared to non-malignant myoblasts. Moreover, the inhibition of calpains using chemical inhibitors led to a decrease in Prx IV abundance (64.32 ± 8.25 and 76.79 ± 4.60 for the precursor and secretable forms, respectively, with calpain inhibitor III treatment). It is the first time that a Prx IV calpain-dependent up-regulation is revealed. In summary, calpains may be implied in the tumour phenotype of ARMS cells especially through Prx IV regulation and may, thus, represent a potential therapeutic target to stop progression of ARMS tumour.