AKR1B10通过靶向糖酵解途径对肝癌细胞增殖和凋亡的影响

Effect of AKR1B10 on Proliferation and Apoptosis of Hepatocellular Carcinoma Cells through Glycolysis Pathway

目的探索醛酮还原酶家族1成员B10(aldo-keto reductase family 1 member B10,AKR1B10)在肝细胞癌(hepatocellular carcinoma,HCC)糖酵解中的功能及其潜在机制。方法利用生物信息学手段分析AKR1B10在HCC中的表达。实时荧光定量聚合酶链反应(real-time reverse transcription PCR,qRT-PCR)和蛋白质印迹(Western blotting)检测AKR1B10的表达;细胞计数试剂盒8(cell counting kit-8,CCK-8)、Transwell实验和流式细胞术分别检测细胞增殖、迁移侵袭和凋亡;qRT-PCR检测骨髓细胞瘤病毒癌基因(myelocytomatosis,MYC)、己糖激酶2(hexokinase 2,HK2)、磷酸甘油酸激酶1(phosphoglycerate kinase 1,PGK1)的表达水平;Seahorse XP96分析胞外酸化率(extracellular acidification rate,ECAR)和耗氧率(oxygen consumption rate,OCR);利用试剂盒检测丙酮酸、乳酸、柠檬酸、苹果酸的含量。结果生物信息学分析结果显示HCC组织和细胞中AKR1B10存在高表达,且GSEA通路富集分析显示其在糖酵解通路上富集。细胞实验发现过表达AKR1B10可以促进HCC细胞增殖、迁移、侵袭,并抑制细胞凋亡。此外,过表达AKR1B10可以促进HCC细胞中MYC、HK2和PGK1的表达,并提高糖酵解水平。回复实验显示糖酵解抑制剂(2-deoxy-D-glucose,2-DG)可以逆转AKR1B10对HCC细胞的增殖、迁移和侵袭的促进作用。结论AKR1B10可以通过激活糖酵解通路促进HCC增殖,提示AKR1B10可能是一种新的HCC诊断标志物和治疗靶点。

Objective Aldo-keto reductase family 1 member B10(AKR1B10)plays an important role in the proliferation and metastasis of malignant tumors including hepatocellular carcinoma(HCC).However,the role of AKR1B10 in HCC glycolysis is not fully understood.Therefore,this study aims to explore the function of AKR1B10 in HCC glycolysis and investigate its potential mechanism.Methods The expression of AKR1B10 in HCC was analyzed by bioinformatics.The expression of AKR1B10 was detected by Real-time reverse transcription PCR(qRT-PCR)and Western blotting.Cell Counting Kit 8(CCK-8),Transwell assay and flow cytometry were used to detect the cell proliferation,migration,invasion and apoptosis,respectively.The expression levels of Myelocytomatosis(MYC),hexokinase 2(HK2)and phosphoglycerate kinase 1(PGK1)were detected by qRT-PCR.Seahorse XP96 was used to analyze the extracellular acidification rate(ECAR)and oxygen consumption rate(OCR).The levels of pyruvate,lactic acid,citric acid and malic acid were detected by kits.Results The bioinformatic analysis showed that AKR1B10 was highly expressed in HCC tissues and cells,and GSEA analysis showed that glycolytic pathway was enriched.Overexpression of AKR1B10 could promote the proliferation,migration and invasion of HCC cells,and inhibit the cell apoptosis.In addition,the overexpression of AKR1B10 could promote the expression of MYC,HK2 and PGK1 in HCC cells,and improve the level of glycolysis.Glycolysis inhibitors(2-Deoxy-D-Glucose,2-DG)could reverse the effect of AKR1B10 on the proliferation,migration,and invasion in HCC cells.Conclusion AKR1B10 can promote the HCC proliferation by activating glycolytic pathway,suggesting that AKR1B10 may be a new diagnostic marker and therapeutic target for HCC.