Cannabis roots have been used in folk medicine for millennia and as nutrient storage systems, contain compounds that may have medicinal value. Despite this, cannabis roots have generally received little attention compared to cannabis flowers, leaves and seeds and were for a long time considered just a waste product. In this paper, for the first time, the extracts of dried roots and inflorescences of Cannabis sativa L. cv Eletta Campana, were chemically investigated in order to compare their metabolite content. The obtained results highlighted a profile rich in fatty acids in the roots and cannabinoids in the inflorescences. Other components such as monoterpenes, sesquiterpenes, diterpenes and triterpenes were also detected. The extracts were also evaluated in terms of cytotoxic activity by using a panel of cancer cell lines derived from different histotypes including melanoma (A375, M14), colon (HCT116, HT29), breast (MDAMB231, MCF7) and non- small cell lung cancer (H1299, A549). Although both extracts significantly reduced the cancer cell viability, the inflorescence extract was more potent. Furthermore, the latter induced a comparable response in all tested cancer cell lines, while melanoma and non-small cell lung cancer were the most sensitive histotypes to the root extract treatment.