Synergistic inhibition of colorectal cancer progression by silencing Aurora A and the targeting protein for Xklp2

To compare effects of AURKA and TPX2 and their combined knockdown on CRC cells.

Unraveling the pathogenesis of colorectal cancer (CRC) can aid in developing prevention and treatment strategies. Aurora kinase A (AURKA) is a key participant in mitotic control and interacts with its co-activator, the targeting protein for Xklp2 (TPX2) microtubule nucleation factor. AURKA is associated with poor clinical outcomes and high risks of CRC recurrence. AURKA/TPX2 co-overexpression in cancer may contribute to tumorigenesis. Despite its pivotal role in CRC development and progression, the action mechanism of AURKA remains unclear. Further research is needed to explore the complex interplay between AURKA and TPX2 and to develop effective targeted treatments for patients with CRC.

Combined knockdown of AURKA and TPX2 was effective in suppressing the malignant phenotype in CRC. Co-inhibition of gene expression is a potential developmental direction for CRC treatment.

We evaluated three CRC gene datasets about CRC (GSE32323, GSE25071, and GSE21510). Potential hub genes associated with CRC onset were identified using the Venn, search tool for the retrieval of interacting genes, and KOBAS platforms, with AURKA and TPX2 emerging as significant factors. Subsequently, cell models with knockdown of AURKA, TPX2, or both were constructed using SW480 and LOVO cells. Quantitative real-time polymerase chain reaction, western blotting, cell counting kit-8, cell cloning assays, flow cytometry, and Transwell assays were used.

Forty-three highly expressed genes and 39 poorly expressed genes overlapped in cancer tissues compared to controls from three datasets. In the protein-protein interaction network of highly expressed genes, AURKA was one of key genes. Its combined score with TPX2 was 0.999, and their co-expression score was 0.846. In CRC cells, knockdown of AURKA, TPX2, or both reduced cell viability and colony number, while blocking G0/G1 phase and enhancing cell apoptosis. Additionally, they were weakened cell proliferation and migration abilities. Furthermore, the expression levels of B-cell lymphoma-2-Associated X, caspase 3, and tumor protein P53, and E-cadherin increased with a decrease in B-cell lymphoma-2, N-cadherin, and vimentin proteins. These effects were amplified when both AURKA and TPX2 were concurrently downregulated.