The thickness shear mode (TSM) resonator attached with living cells has been shown to be an effective functional biosensing device to monitor the process of cell adhesion to a surface. In this study, we first monitored the dynamic process of cell attachment and spreading as a function of cell seeding densities. Based on the steady state of cell adhesion to the substrate, a multilayer sensor structure model including a quartz substrate, a cell-substrate interfacial layer and a cell layer was constructed. The thickness of cell-substrate interfacial layer and the viscoelastic properties of human skin fibroblasts (HSF) were then determined by fitting experimental results with the theoretical model. It has been obtained that the thickness of the cell-substrate interfacial layer is 60-80 nm, and the elastic module and viscosity of cell layers are about 13 KPa and 3-4 mPa's respectively. These results are in a good agreement with those measured by other techniques, such as magnetic bead microrheometry, atomic force microscopy (AFM) and Surface Plasmon Resonance Microscopy (SPRM). In addition, knowing that the actin cytoskeleton is important for the mechanical properties of living cells, we investigated the motional resistance change caused by the disruption of actin cytoskeleton induced by fungal toxin Cytochalasin D in the human skin fibroblasts. The results indeed indicate the direct correlation between resistance changes and the disruption of actin cytoskeleton, which are again consistent with the results observed by fluorescence images.