Lichens are a symbiotic relationship between a fungus and a photosynthetic partner. Chemical characterization and bioactive potentials (antiproliferative, antioxidant, and antibacterial) of five lichen species (Evernia prunastri, Platismatia glauca, Pseudevernia furfuracea, Ramalina fastigiata, and Ramalina farinacea) were assessed. Five lichen metabolites (usnic acid, atranorin, stictic acid, evernic acid, and fumarprotocetraric acid) were analyzed by HPLC-DAD. E. prunastri was noteworthy evernic acid source. Antiproliferative activity was evaluated using human breast adenocarcinoma (MCF-7) and human hepatocellular carcinoma (HepG2/C3A) cell lines. The strongest activity was observed for P. glauca against HepG2/C3A, while the only lichen species that induced cytotoxicity against MCF-7 cell line was P. furfuracea. The highest antioxidant activity was also obtained with P. furfuracea. E. prunastri and R. farinaceae had the highest phenolic and flavonoid contents, respectively. Antibacterial activities of the extracts were determined against ten pathogenic bacteria. The most effective antibacterial agent was methanol extract of R. fastigiata. Our findings have revealed the pharmaceutical potentials of tested lichen species.