Even though the outdoor air pollution and its major component Particulate Matter (PM) are recently classified as human carcinogen, attempts to elucidate the underlying mechanisms of PM toxicity are still crucial and continuing with in vitro approaches in various environmental circumstances. Present study investigated the genotoxicity (Comet assay) and the cytotoxicity (lactate dehydrogenase (LDH) leakage and the water-soluble tetrazolium (WST-1) assays) of 30 daily PM2.5 samples collected in the Kütahya province, to address their daily variability in effects with season (i.e. winter versus summer) and location (i.e. rural versus urban) using A549 human lung cancer epithelial cell line, as well as in relation to their chemical composition, specifically trace elements, organic carbon (OC) and elemental carbon (EC). The genotoxicity, measured by the percentage tail intensity (TI), of the daily PM2.5 samples at the traffic dense urban station was higher than that of the rural site for 80% of the parallel days. The genotoxicity was significant in the winter at the urban and in the summer at the rural site. Cytotoxicity was the highest for the winter urban samples. The PM2.5 mass, OC, and EC concentrations were not correlated to DNA damage, while there were correlations with Mn, Fe, Cu and Ba at the rural PM2.5 samples, and Mn, Co and Ni at the urban samples, respectively. The present study is confirming that the complex composition of PM2.5 originating from spatial and temporal changes can cause differences in the health effects.