Knock‐down of plasminogen‐activator inhibitor‐1 enhances expression of E‐cadherin and promotes epithelial differentiation of human pancreatic adenocarcinoma cells

Abstract

High levels of plasminogen activator inhibitor-1 (PAI-1), which is produced by stromal, endothelial and cancer cells and has multiple complex effects on cancers, correlate with poor cancer prognosis. To more definitively study the role of endogenously produced PAI-1 in human pancreatic adenocarcinoma (PAC) PANC-1 cell line biology, we used anti-PAI-1 shRNA to create stable PAI-1 deficient cells (PD-PANC-1s). PD-PANC-1s exhibited a heterogeneous morphology. While the majority of cells exhibited a cuboidal shape similar to the parental PANC-1 or the vector-infected control cells, numerous large cells with long filopodia and a neuronal-like appearance were observed. Although both Vector-control cells and PD-PANC-1s expressed mRNAs that are characteristic of mesenchymal, neural and epithelial phenotypes, epithelial marker RNAs were up-regulated (e.g. E-cadherin, 32-fold) whereas mesenchymal marker RNAs were down-regulated (e.g. Thy1, 9-fold) in PD-PANC-1s, suggesting mesenchymal-to-epithelial transition. Neural markers exhibited both up- and down-regulation. Immunocytochemistry indicated that epithelial-like PD-PANC-1s expressed E-cadherin and β-catenin in significantly more cells, while neural-like cells exhibited robust expression of organized β-3-tubulin. PAI-1 and E-cadherin were rarely co-expressed in the same cells. Indeed, examination of PAI-1 and E-cadherin mRNAs expression in additional cell lines yielded clear inverse correlation. Indeed, infection of Colo357 PAC cells (that exhibit high expression of E-cadherin) with PAI-1-expressing adenovirus led to a marked decrease in E-cadherin expression and to enhanced migration of cells from clusters. Our results suggest that endogenous PAI-1 suppresses expression of E-cadherin and differentiation in PAC cells in vitro, supporting its negative impact on tumor prognosis.

Immunohistochemical expression of NEDD9, E-cadherin and γ-catenin and their prognostic significance in pancreatic ductal adenocarcinoma (PDAC)

“…Focally lost E-cadherin expression was more common in our sample and it was also correlated with intermediate γ-catenin expression. Lupu-Meiri et al suggested plasminogen activator inhibitor-1 (PAI-1) as another factor that suppresses differentiation of PDAC cells by inhibiting E-cadherin expression [24].…”

Early Epigenetic Downregulation of microRNA-192 Expression Promotes Pancreatic Cancer Progression

“…In advanced pancreatic cancer, PAI-1 levels predict response to gemcitabine chemotherapy (38) and pharmacological inhibition of uPA in combination with gemcitabine is a promising new treatment option for pancreatic cancer (39). Additionally, overexpression of PAI-1 is a prominent feature of EMT (40). Therefore, upregulation of PAI-1 together with EMT resulting from low miR-192 levels could trigger cell migration and contribute to an invasive phenotype.…”

Cancer-Associated Thrombosis: An Overview of Mechanisms, Risk Factors, and Treatment

“…PAI-1 is a key inhibitor of fibrinolysis which has been shown to be highly expressed in pancreatic cancer cells [ 89 ]. Increased PAI-1 in plasma results in reduced fibrinolytic activity, thus increasing the risk of thrombosis [ 90 ].…”