This study investigates cytotoxic and genotoxic activities of (+)-Usnic acid and (-)-usnic acid isolated from the lichen Ramalina farinacea and the lichen Cladonia foliacea, respectively. To determine the activities of these acids, we used the MTT assay on V79 (Chinese hamster lung fibroblast like) and A549 (human lung carcinoma epithelial like) cell lines and cytokinesis-blocked micronucleus (CBMN) assay in human lymphocytes in vitro. Our results suggest that both enantiomers of usnic acid are non-genotoxic shown by the absence of micronucleus induction in human lymphocytes and have significant cytotoxic and apoptotic effects to induce cell killing in cultured human lymphocytes, V79 and A549 cell lines. Even low doses of (+)-usnic acid showed high cytotoxic activity against cancerous cells. The MTT results and cell proliferation index (CPI) values based on the CBMN test results are found in good agreement.