Study on the Mechanism of SIRT2 Regulating Aerobic Glycolysis and Growth Proliferation of Colorectal Cancer Cells

LI Zhiran; DONG Yujia; ZHANG Ruonan; LIU Sitong; YUE Jinjin

doi:10.3969/j.issn.2096-305X.2025.06.003

Abstract：Objective To explore the biological function of the silent information regulator factor 2(SIRT2)in the metabolic reprogramming and proliferation regulation of colorectal cancer cells,as well as its impact on the glucose metabolism pathway,At the same time to study the protein expression levels of SIRT2 and hexokinase Ⅱ(HK Ⅱ)in colorectal cancer and their correlation.Methods Western Blot was used to analyze the protein expression level of SIRT2 in colorectal cancer cells.Based on the results,cell lines with high SIRT2 expression were selected for gene interference,while cell lines with low SIRT2 expression were used for overexpression studies.The glucose concentration in the culture medium was measured using the glucose oxidase method,and lactate levels were analyzed colorimetrically to study the metabolic state of the cells.Cell viability was detected by CCK-8 assay,and proliferative capacity was assessed by colony formation assay.Additionally,reverse transcription PCR(RT-PCR)was used to study the effect of SIRT2 interference or overexpression on HK Ⅱ mRNA expression levels,and Western Blot was performed to further validate the effect of SIRT2 regulation on HK Ⅱ protein expression.Immunohistochemistry was used to detect the protein expression levels of SIRT2 and HK Ⅱ in colorectal cancer tissues.Rescue experiments were conducted by co-transfecting SIRT2-siRNA and HK Ⅱplasmids,followed by measurements of glucose and lactate levels in the culture medium,as well as CCK-8 and colony formation assays for each group.Results After RNA interference,compared with the negative control group,the interference group exhibited suppressed glycolysis,characterized by increased glucose levels and decreased lactate production,along with significantly reduced cell proliferation activity and colony formation ability.Both HK Ⅱ mRNA and protein expression levels decreased.After overexpression experiments,compared with the control group,overexpression significantly enhanced aerobic glycolysis in colorectal cancer cells,manifested as increased glucose consumption and lactate accumulation in the culture medium,while also promoting cell growth,proliferation,and colony formation efficiency.Both HK Ⅱ mRNA and protein expression levels increased.Immunohistochemistry results showed that the expression levels of SIRT2 and HK Ⅱ in colorectal cancer tissues were significantly higher than those in paired adjacent tissues.In the rescue experiments,compared with the interference group,the co-transfection group showed significantly decreased glucose levels,significantly increased lactate levels,and markedly enhanced cell proliferation activity and colony formation rate.Conclusion SIRT2 promotes aerobic glycolysis and cell growth/proliferation in colorectal cancer cells,and the mechanism may be related to SIRT2 regulating HK Ⅱ gene expression.