The PIEZO1 Knockout hTERT-HPNE Cell Line is a CRISPR/Cas9-edited knockout cell line that provides a loss-of-function model for PIEZO1, encoding a mechanosensitive cation channel. This engineered cell line enables targeted disruption of the PIEZO1 gene, facilitating investigations into mechanotransduction and calcium signaling in a pancreatic ductal epithelial context. The knockout model is suited for studying PIEZO1-dependent processes such as cell volume regulation, migration, and differentiation. Researchers can interrogate PIEZO1 function in a controlled in vitro system without interference from endogenous channel activity.
The parental hTERT-HPNE cell line is an immortalized normal human pancreatic ductal epithelial cell model. These cells were derived from primary pancreatic ductal epithelium and immortalized via hTERT expression, retaining key features of their primary counterparts. hTERT-HPNE cells maintain the ability to secrete bicarbonate-rich fluid and form monolayers that recapitulate the pancreatic ductal barrier. This background makes them a physiologically relevant platform for exploring ductal cell biology, pancreatitis, and early neoplastic transformation in the pancreas.
PIEZO1 functions as a mechanosensitive cation channel that opens in response to membrane tension, shear stress, or osmotic stimuli, permitting calcium influx. This calcium signal is transduced through calpain, calcineurin, and NFAT, while also engaging integrin-mediated pathways and the MAPK/ERK cascade. Interacting partners such as STOML3 and PACSIN2 modulate channel sensitivity and trafficking, and cytoskeletal proteins link PIEZO1 to cellular mechanics. Downstream, calcium influx activates FAK and Src, promoting ERK1/2 phosphorylation and YAP/TAZ nuclear translocation, thereby integrating physical stimuli with transcriptional programs that control proliferation and migration.
In pancreatic ductal epithelium, PIEZO1-mediated calcium signaling regulates critical functions including epithelial integrity, ductal fluid secretion, and cell migration. Aberrant PIEZO1 activity is implicated in pancreatic cancer progression, where it may promote tumor cell migration and epithelial-mesenchymal transition. Use of this knockout cell line in the hTERT-HPNE background permits dissection of PIEZO1??s contributions to pancreatic ductal cell pathophysiology. It is particularly relevant for studying mechanotransduction in pancreatitis-associated fibrosis and for evaluating PIEZO1 as a target in pancreatic ductal adenocarcinoma initiation.
Typical applications include mechanotransduction studies in pancreatic epithelium, investigation of pancreatic cancer initiation and progression, pancreatitis research, and ductal cell function assays. Researchers can employ calcium imaging and patch-clamp electrophysiology to characterize loss of mechanosensitive currents, cell migration assays to assess PIEZO1-dependent motility, western blotting for phospho-ERK to monitor MAPK pathway activation, and immunofluorescence or flow cytometry to examine proliferation and apoptosis. For additional information or technical inquiries, please contact Ascent Research.
