Expression of Akt3 in colorectal cancer and its effect on biological behavior of colorectal cancer cells

SUN Haiying; JIA Wei; XUE Mei; MA Chao; ZHANG Mengqiao; WANG Jing; SHUAI Junfang; SUN Fanyao

doi:10.3969/j.issn.1009-0460.2025.01.001

Abstract：Objective To investigate the expression of serine/threonine kinase 3(Akt3)in colorectal cancer(CRC)and its impact on the biological functions of CRC cells.Methods The Tumor Genome Atlas(TCGA)database and the Gene Expression Omnibus(GEO)database were utilized to screen out the nestin apoptosis-related genes(ARGs)that were significantly differentially expressed in CRC tissues and affected prognosis.The role of Akt3 in CRC was explored through text mining and functional enrichment analysis.The lentivirus transfection technique was used to overexpress Akt3 in SW480 cells,and the overexpression efficiency was verified by Western blotting.Flow cytometry was used to detect the apoptosis ability of cells,and the scratch healing assay and Transwell invasion assay were used to evaluate the migration and invasion abilities of cells.Results Based on the analysis results from the TCGA and GEO databases,14 ARGs were closely related to CRC and its prognosis were ultimately determined.Bioinformatics analysis revealed that the expression of Akt3 in CRC tissues was significantly lower than that in normal tissues,and the survival rate of patients with low expression of Akt3 was significantly lower than that of the high expression group(P=0.009).The results of GO functional enrichment analysis indicated that Akt3-related genes were mainly enriched in extracellular matrix organization,extracellular structure organization,external encapsulating structure organization,collagen fiber organization,and cell-matrix adhesion,among other cellular components and biological processes.The results of KEGG pathway enrichment analysis showed that Akt3-related pathways were mainly concentrated in the cytoskeleton of muscle cells,cell adhesion molecules,staphylococcus aureus infection,complement and coagulation cascades,and extracellular matrix-receptor interaction signaling pathways.Western blotting experiments demonstrated that the expression level of Akt3 in the overexpression plasmid/slow virus group was significantly higher than that in the control group(P=0.002).Flow cytometry detection showed that the apoptosis rate of SW480 cells in the Akt3 overexpression group was significantly increased(P=0.004).Transwell invasion experiments indicated that the number of cells that penetrated the upper chamber matrix gel was significantly reduced after the overexpression of Akt3(P=0.012).The results of the scratch healing experiment showed that the average healing rate of the Akt3 overexpression group was 52.0%,significantly slower than 74.2%of the control group(P=0.005).Conclusion Akt3 is underexpressed in colorectal cancer and its expression level is significantly correlated with patients′ prognosis.Akt3 may serve as a potential biomarker and therapeutic target for CRC.