Panoply™ Human YAP1 Knockdown Stable Cell Line

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies and carries an extremely high mortality rate. Recent studies have demonstrated that metabolic alterations play a crucial role in maintaining the malignant phenotype of pancreatic cancer; however, the molecular mechanisms underlying glucose metabolic reprogramming remain unclear. This study aimed to elucidate the role of Yes-associated protein (YAP1)-a key effector of the Hippo pathway-in regulating aerobic glycolysis in pancreatic cancer. Researchers found that silencing YAP1 expression inhibited anabolic glycolysis in pancreatic cancer cells. YAP1 was shown to regulate HIF-1α protein levels, transcriptional activity, and the expression of HIF-1α-targeted glycolytic genes. Further analysis revealed that EGLN2, a regulator of HIF-1α protein levels, is a direct target of YAP1. Low EGLN2 expression was associated with poor patient prognosis. Analyses of the TCGA dataset and immunohistochemical staining confirmed an inverse correlation between YAP1 and EGLN2 expression at both the mRNA and protein levels. These findings suggest that YAP1 holds promise as a predictive biomarker and therapeutic target for human pancreatic cancer.

YAP1 positively regulates glycolysis in pancreatic cancer. To investigate the underlying mechanism, researchers examined HIF-1α protein levels in YAP1-knockdown PANC-1 and MIA PaCa-2 cells. HIF-1α is a regulator of anabolic glycolysis that enables cancer cells to adapt to hypoxic environments. The results showed that YAP1 knockdown led to a significant reduction in HIF-1α protein levels (Figure 1A). Overexpression of HIF-1α in YAP1-knockdown PANC-1 cells increased the extracellular acidification rate (ECAR) and correspondingly decreased the oxygen consumption rate (OCR), indicating that YAP1 regulates glycolysis in pancreatic ductal adenocarcinoma (PDAC) via HIF-1α (Figure 1B, C). Next, the researchers determined the regulatory effect of YAP1 on HIF-1α transcriptional activity using a dual-luciferase reporter assay. HRE-luciferase reporter assay results demonstrated that YAP1 positively regulates HIF-1α transcriptional activity in a dose-dependent manner (Figure 1D). HIF-1α regulates glycolysis by transcriptionally activating a series of glycolysis-related genes; notably, GLUT1, HK2, and LDHA are well-established HIF-1α target genes involved in glycolysis. Subsequently, the researchers examined the expression of these glycolytic genes in YAP1-knockdown PANC-1 and MIA PaCa-2 cells. The results showed that reduced YAP1 expression led to a corresponding downregulation of GLUT1, HK2, and LDHA expression (Figure 1E). Consistent with the quantitative PCR results, the protein levels of these glycolytic genes also decreased in YAP1-knockdown cells, further confirming the positive role of YAP1 in regulating HIF-1α protein levels and transcriptional activity (Figure 1F).

Figure 1. YAP1 regulated HIF‐1α and HIF‐1α transcriptional activity in pancreatic cancer. (Hu P, et al., 2024)