Abstract Chemical composition of essential oil (EO) and methanol extracts (MEs) from different parts of Tanacetum vulgare L. plant was analyzed and investigated for potential biological activities and correlated with the main constituents detected in EO and MEs. The EO was characterized by a high content of oxygenated monoterpenes with trans-chrysanthenyl acetate as major compound. All MEs were characterized by neochlorogenic, 3,5-O-dicaffeoylquinic and caffeoylquinic acids. High phenolic content in MEs correlated to high antioxidant capacity, especially for roots. Tansy EO showed strong activity against the most of tested fungi and was efficient as bifonazole and ketoconazole, while the most sensitive bacteria were Gram-negative E. coli and E. cloacae. All MEs showed fungistatic and fungicidal effects against all of the eight tested fungi, but the antimicrobial activity was higher against Gram-positive bacteria. Additionally, shown for the first time, MEs of leaves and flowers exhibited a strong antiproliferative effect on human cervical adenocarcinoma (HeLa) cells, causing cell shrinkage and detachment. In the present study, the Tansy extracts and essential oil with low thujone content can provide a very promising and effective alternative in the field of antimicrobial applications and food preservation. The Tansy MEs possess a high antioxidant potential with phenolic acids being a major radical scavenging contributor with the proved antiproliferative activity to HeLa cells.