Recently microRNA-26a(miR-26a)has been revealed to perform manifold biological functions in different cancers,including control of cellular differentiation,cell growth,apoptosis,metastasis and cell metabolism.In our study,we focus on exploring miR-26a functions in glucose metabolism of colorectal cancer(CRC)cells using a SILAC-based proteomic strategy.A total of 4833 proteins were quantified in the miR-26a overexpressing CRC cells,of which 437 were significantly altered(fold change > 1.5 and p-value < 0.01).Gene Ontology analysis indicated that almost half of the differential expressed proteins take part in cell metabolism process,including insulin carbon metabolism,butanoate metabolism,biosynthesis of amino acids and glycerolipid metabolism pathways.Except for some key enzymes in metabolism,such as HKDC1,ALDH1A3,ME1,G6PD,IDH1,MLYCD,and ACO2,several other candidates were also identified to change their expression levels.For example,TIGAR(fructose-2,6-bisphosphatase),which was 1.86-fold decreased,potentially promotes the pentose phosphate pathway to produce NADPH for antioxidant function and ribose-5-phosphate for nucleotide synthesis in intestinal cancer.Furthermore,we discovered that miR-26a direct targets the 3-UTR of PDHX mRNA to inhibit the expression of PDHX,which inhibits the conversion of pyruvate to acetyl coenzyme A in the citric acid cycle.And the overexpression of miR-26a in HCT116 cells efficiently improved the accumulation of pyruvate and decreased the production of acetyl coenzyme A.Meanwhile the inhibition of miR-26a expression induced inverse biological effects.Taken together,miR-26a regulates glucose metabolism of colorectal cancer cells by affecting multiple metabolism enzymes/proteins,especially directly inhibiting PDHX to contribute to aerobic glycolysis in CRC cells.