A prospecting search for antitumoural activity in polar benthic invertebrates was conducted on Antarctic and sub-Antarctic benthos in three different areas: Bouvet Island (sub-Antarctic), eastern Weddell Sea (Antarctica) and the South Shetland Islands (Antarctica). A total of 770 benthic invertebrate samples (corresponding to at least 290 different species) from 12 different phyla were assayed to establish their pharmacological potential against three human tumour cell lines (colorectal adenocarcinoma, lung carcinoma and breast adenocarcinoma). Bioassays resulted in 15 different species showing anticancer activity corresponding to five different phyla: Tunicata (5), Porifera (4), Cnidaria (3), Echinodermata (2) and Annelida (1). This appears to be the largest pharmacological study ever carried out in Antarctica and it shows very promising antitumoural activities in the Antarctic and sub-Antarctic benthos.