Investigation of the anti-metastatic role of flavopiridol via epithelial-mesenchymal transition in pancreatic ductal adenocarcinoma cells

Pancreatic cancer is among the most lethal malignancies worldwide, characterized by an exceptionally low survival rate. The aggressive nature of the disease, marked by its high metastatic potential and the limitations in current diagnostic and therapeutic strategies, results in rapid disease progression and poor patient outcomes. Consequently, extensive research efforts are underway to develop more effective treatment modalities. In this study, the effects of Flavopiridol (FP), a synthetic flavonoid, on epithelial-mesenchymal transition (EMT) were investigated in pancreatic cancer cell lines Panc-1 and MIA PaCa-2. The experimental findings demonstrated that FP exhibited the most potent effects at concentrations of 300, 400, and 500 nM following 24 hours of treatment and at 100, 200, and 300 nM after 48 hours of exposure. The impact of FP on EMT markers was evaluated at the protein level. In Panc-1 cells, the most significant reduction in the expression of ZEB1 and Vimentin was observed with 300 nM FP treatment for 48 hours. In MIA PaCa-2 cells, a marked decrease in ZEB1 expression was detected after 48 hours of treatment with 300 nM FP, while Vimentin expression was significantly downregulated following both 24-hour treatment with 500 nM FP and 48-hour treatment with 300 nM FP. Moreover, a notable reduction in the expression levels of Snail and N-Cadherin was observed across all FP concentrations after 48 hours of treatment. Interestingly, FP treatment resulted in the upregulation of the epithelial markers Claudin and E-Cadherin at the protein level, particularly following 24 hours of exposure. At the transcriptional level, in Panc-1 cells, Claudin mRNA expression was significantly increased with 400 nM FP treatment for 24 hours and 300 nM FP treatment for 48 hours. Additionally, E-Cadherin mRNA expression was upregulated at both time points with 300 nM FP treatment. Conversely, Vimentin mRNA expression was downregulated with 300 nM FP treatment at both 24 and 48 hours, while ZEB1 mRNA expression was significantly reduced following 300 nM FP treatment for 24 hours and 200 nM FP treatment for 48 hours. In MIA PaCa-2 cells, Claudin mRNA expression increased with 500 nM FP treatment for 24 hours and 200 nM FP for 48 hours, whereas E-Cadherin mRNA levels were elevated following 500 nM FP treatment for 24 hours and 300 nM FP for 48 hours. Furthermore, Vimentin mRNA expression was consistently downregulated with 300 nM FP treatment at both time points, while ZEB1 mRNA expression was significantly decreased following 300 nM FP treatment for 24 hours and 200 nM FP for 48 hours. These findings suggest that Flavopiridol exerts a significant modulatory effect on EMT-associated markers in pancreatic cancer cells, potentially contributing to the inhibition of metastatic progression. Comprehensive analyses have demonstrated that FP significantly upregulates the expression of epithelial mediators while downregulating the expression of mesenchymal mediators in pancreatic cancer cells.