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Overexpression pattern and molecular biological mechanism of RAD21 in pancreatic adenocarcinoma: a study integrating bulk RNA-Seq, scRNA-Seq, stRNA-Seq, CRISPR screening and immunohistochemistry

bioinformatics
molecular-biology
histology

Abstract

Background. RAD21 significantly contributes to the emergence and progression of numerous aggressive cancers. However, the specific role of RAD21 in pancreatic adenocarcinoma (PAAD) remains unknown. Materials and methods. In this research, for the first time, 2044 global mRNA expression datasets, complemented by 64 in-house tissues of immunohistochemistry analysis, were utilised to assess the irregular expression patterns of RAD21 in PAAD. Clustered regularly interspaced short palindromic repeats knockout screening technology was used to evaluate the necessity of RAD21 for the growth of 25 PAAD cell lines. The metabolic activity, inferCNV and WGCNA of the scRNA-Seq data, deconvolution algorithm and cellChat analysis of the stRNA-Seq data were used to further explore the potential molecular biological mechanisms of RAD21 in PAAD. Receiver operating characteristic (ROC) curves, summary receiver operating characteristic (sROC) curves, positive likelihood ratios and negative likelihood ratio forest plots were used to evaluate the ability of RAD21 to predict PAAD. Results. RAD21 mRNA was significantly overexpressed in PAAD tissues (SMD = 0.76, 95% CI 0.36–1.16, p<0.05). Verification at the protein level also demonstrated the high expression of RAD21 in PAAD tissues (p<0.05). The growth of PAAD cell lines was significantly inhibited after RAD21 was knocked out. Functional enrichment analysis revealed that RAD21 in PAAD was related to cell cycle-related pathways. ScRNA-Seq analysis revealed that RAD21 may be related to the growth of PAAD malignant epithelial cells. RNA-Seq analysis suggested that RAD21 in malignant epithelial cells may act on the WNT signalling pathway. In addition, the expression levels of RAD21 mRNA (AUC=0.83, 95% CI=0.79–0.86) and protein (AUC=0.865) strongly affected PAAD tissues. Conclusion. RAD21 has a significantly high expression pattern in PAAD tissues. RAD21 may affect the growth of PAAD cells by acting on the WNT-related pathway.
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