Pancreatic ductal adenocarcinoma (PDAC) is the fourth most common leading cause of cancer-related deaths worldwide and is characterized by the formation of a dense fibrotic stroma. The excessive amount of extracellular matrix (ECM) is primarily formed by pancreatic stellate cells (PSCs). ECM contributes to a high pancreatic tissue pressure which itself also stimulates PSCs. The effect of increasing tissue pressure and altered biomechanics in the pathogenesis of PDAC, however, is poorly understood. Mechano-sensing and -transduction have been linked to opening of mechanosensitive ion channels such as PIEZO1, TRPV4 or TRPC1. We hypothesize that these channels whose expression we showed in PSCs, play crucial roles in sensing/transducing mechanical cues that are typical for the PDAC microenvironment. We have shown that TRPC1 channels participate in the activation of mechanically stressed PSCs. TRPC1 knock-out is sufficient to attenuate the pressure-induced activation (Fels et al. 2016 Eur Biophys). We performed live-cell imaging migration experiments and monitored Ca2+ influx into PSCs by means of the Mn2+ quench technique in order to assess the function of PIEZO1 channels in PSCs. Stimulation of PIEZO1 channels with its activator Yoda1 leads to a doubling of the translocation of migrating PSCs (41.2 ± 4.3 µm vs. 20.4 ± 2.2 µm) and enhanced directionality by 67 ± 10 %. Activation of PIEZO1 leads to a fourfold increase of calcium influx into the PSCs. We then focused on the PIEZO1-mediated effect on the 3D invasion of PSCs. Cell spheroids were embedded in a 3D desmoplastic matrix and invasion was analyzed for a time period of 48h with or without PIEZO1 stimulation. After 24h, invasion area of Yoda1-stimulated PSCs is 10% lower than that of control cells. However, PIEZO1 activation causes the number of detached cells from the spheroid core to increase by 68%. Additionally, upon PIEZO1 activation PSCs transmit more force to the surrounding ECM, as revealed by measuring the dislocation of microbeads embedded in the surrounding matrix (control: 9.0 ± 0.4 µm/h; Yoda1: 11.3 ± 0.4 µm/h). In summary, stimulating PIEZO1 activates PSCs by inducing calcium influx. This results in increased motility within the ECM. Activated PSCs thereby transfer more force to the matrix, possibly align ECM fibers and subsequently improve their invasion capacity and detach from the primary spheroid. We hypothesize a strong interdependence between the mechanical output of PSCs and stromal mechanics, which promotes early local invasion of PDAC cells.