Molecular mechanisms of RACK1-driven metastasis in pancreatic ductal adenocarcinoma revealed by single-cell RNA sequencing.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with significant cellular heterogeneity, which poses a significant challenge for treatment. We integrated three single-cell RNA sequencing (scRNA-seq) datasets to construct an expression matrix, identify cell types, and assess copy number variation (CNV) scores, with a particular focus on ductal cells. Our study found that ductal cells are the primary source of malignant cells in PDAC and identified a high-malignancy subtype—ductal cell-3—where RACK1 was the most significantly differentially expressed gene. Through the analysis of public and clinical data, along with RT-qPCR and immunohistochemistry validation, we confirmed the high expression of RACK1 in PDAC tumors, which correlated with lymph node metastasis, clinical staging, histological differentiation, and prognosis. Further investigation revealed that RACK1 promotes PDAC cell proliferation, migration, and invasion. Sequencing analysis indicated that RACK1 inhibits the FOXO signaling pathway, promotes epithelial-mesenchymal transition (EMT), and facilitates the progression and metastasis of PDAC. This study provides new insights into the role of RACK1 in PDAC and its underlying mechanisms.