SPAG4 Regulates Glycolytic Metabolism in HT29 Cells as a Target via the c-MYC/SULT2B1 Pathway

Glycolytic metabolism has been identified as a facilitator of tumor cell proliferation. Therefore, this study aims to investigate the mechanisms by which the sperm-associated antigen 4 (SPAG4)/cellular myelocytomatosis oncogene (c-Myc)/sulfotransferase 2B1 (SULT2B1) axis regulates glycolytic metabolism and influences the viability of HT29 cells.

SPAG4, c-Myc, and SULT2B1 levels were assessed in HT29 cells using Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Western blot analyses. Moreover, overexpression and knockdown in HT29 cell models were successfully established. Furthermore, cell viability and proliferation were evaluated using Cell Counting Kit-8 (CCK-8) and colony formation assays. Various key parameters such as glucose uptake, lactate production, Adenosine Triphosphate (ATP)/Adenosine Diphosphate (ADP) ratio, and the expression levels of Glucose transporter 1 (GLUT1) and Lactate dehydrogenase A (LDHA) were determined to examine glycolytic metabolism. Additionally, the relationship between SPAG4, c-Myc, SULT2B1, and glycolysis was assessed using the immunofluorescence staining approach and 2-Deoxy-D-glucose (2-DG) therapy.

The expression levels of SPAG4, c-Myc, and SULT2B1 were significantly elevated in HT29 cells (p < 0.05). Moreover, silencing SPAG4 and c-Myc substantially reduced glycolytic metabolism and suppressed HT29 cell viability and colony formation capability (p < 0.05). Additionally, elevated SULT2B1 expression effectively counteracted the glycolytic reduction induced by silencing SPAG4 and c-Myc, enhancing cellular viability and colony formation capability (p < 0.05).

In summary, SPAG4 knockdown effectively suppresses HT29 cell proliferation and colony formation ability by decreasing SULT2B1 expression through the downregulation of c-Myc, leading to the reduction of glycolytic metabolism.