Abstract

Introduction: Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide, with metastasis significantly reducing patient survival. Despite advances in treatment, the molecular mechanisms underlying the progression of colorectal cancer remain poorly understood. Recent studies highlight the role of TPX2 and KIF20A, two proteins involved in cell division, in the development of cancer. TPX2 plays a key role in the assembly of the mitotic spindle, while KIF20A is a member of the kinesin superfamily, which is important for intracellular transport and cytokinesis. Both genes are associated with various types of cancer, but their specific contribution to CRC remains unclear. The aim of this study is to investigate the expression and prognostic significance of TPX2 and KIF20A in CRC through bioinformatic analysis and experimental validation. Methods: To identify differentially expressed genes (DEGs) in CRC, five publicly available microarray datasets (GSE39582, GSE8671, GSE9348, GSE21510, and GSE44076) were analyzed using the Limma package in R. Functional enrichment analysis of DEGs was performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. A protein-protein interaction (PPI) network was created using STRING and visualized using Cytoscape to identify hub genes. Survival analysis of hub genes was performed using the Gene Expression Profiling Interactive Analysis (GEPIA) tool with TCGA data. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic potential of hub genes. Experimental validation was performed on 50 CRC samples using quantitative real-time PCR (qRT-PCR) to measure the expression of TPX2 and KIF20A. Results: The integrated analysis identified several DEGs significantly involved in CRC, with TPX2 and KIF20A emerging as key hub genes. GO and KEGG analyses showed that these genes are highly associated with cell cycle regulation and mitotic processes. Survival analysis showed that high TPX2 and KIF20A expression correlates with poorer prognosis in colorectal cancer patients. ROC curve analysis confirmed their potential as diagnostic biomarkers. Experimental validation showed significant upregulation of TPX2 and KIF20A in CRC tissues compared to normal controls, supporting the bioinformatic results. Further mechanistic evidence suggests that TPX2 and KIF20A contribute to colorectal cancer progression by promoting cell proliferation and tumor formation. Previous studies also suggest that KIF20A activates the JAK/STAT3 signaling pathway, thereby increasing the aggressiveness of colorectal cancer cells, while TPX2 is associated with chromosomal instability and tumorigenesis. These results suggest that targeting TPX2 and KIF20A may offer new therapeutic opportunities for the treatment of colorectal cancer. Conclusion: This study highlights the potential role of TPX2 and KIF20A as prognostic biomarkers and therapeutic targets in colorectal cancer. Their significant upregulation in tumor tissue and strong association with poor survival outcomes underscore their importance in colorectal cancer progression. Future research should focus on elucidating the molecular mechanisms underlying their oncogenic role and exploring targeted therapies aimed at modulating their activity to improve outcomes for colorectal cancer patients. © Biomedpress.