PM2.5 is one of the important pollutants, so it is of great significance to the study of cytotoxical effects of PM2.5 from different regions. In this study, Human lung carcinoma epithelial cells (A549) were exposed to the average daily volume of PM2.5 from three different regions in vitro for 2 h. Afterwards, the cells were examined on the single cell level by the confocal Raman microspectroscopy and atomic force microscopy, respectively, to obtain the Raman spectroscopy, and the morphological and biomechanical properties of the cells. In addition, the cytokine concentration was measured by the multiplex enzyme linked immunosorbent assay. Compared to the untreated cells, statistical results revealed several interesting phenomena, including (1) the intensity of the Raman peaks of the treated cells increased at 1 002.97, 1 127.29, 1 172.83 and 1338.1 cm-1; (2) the lamellipodia of treated cells shrunk or partially disappeared; (3) the treated cells became higher; (4) the adhesion work decreased while the elastic energy increased for the cells treated by the PM2.5 from Suzhou region and (5) the treated cells had an inflammatory response. Our work demonstrated that the toxical effects of PM2.5 from different regions on A549 were different, which provides useful information for the air pollution control and prevention and control of lung diseases.