FOXM1转录激活HJURP介导有氧糖酵解促进乳腺癌细胞的阿霉素耐药性

FOXM1 transcriptionally activated HJURP mediated aerobic glycolysis promotes doxorubicin resistance in breast cancer cells

目的本研究旨在阐明乳腺癌中有氧糖酵解调控阿霉素耐药的作用机制。方法生信分析乳腺癌组织中霍利迪连接识别蛋白(HJURP)和叉头核蛋白1(FOXM1)的表达及其相关性,并分析HJURP的富集通路;qPCR检测HJURP和FOXM1在乳腺癌细胞中的表达;及在阿霉素耐药细胞中的表达。双荧光素酶实验和ChIP实验验证FOXM1与HJURP的结合关系;CCK-8检测细胞活力和IC 50值;Western blot检测糖酵解代谢途径相关的特定基因的表达;Seahorse XP96分析不同处理组的胞外酸化率(ECAR)和耗氧率(OCR),试剂盒检测各处理组细胞中丙酮酸、乳酸和ATP水平。结果HJURP在乳腺癌中高表达,富集在糖酵解通路上。细胞功能实验发现HJURP能够通过调节有氧糖酵解促进细胞阿霉素耐药性。此外FOXM1靶向结合HJURP,并在乳腺癌中高表达,与HJURP呈正相关。回复实验发现HJURP过表达能够逆转FOXM1敲低对乳腺癌细胞阿霉素耐药性的抑制作用。结论本研究结果证明转录因子FOXM1能够激活HJURP的转录调节有氧糖酵解促进乳腺癌细胞阿霉素耐药性。

Objective This study aimed to elucidate the mechanism of aerobic glycolysis regulating doxorubicin resistance in breast cancer.Methods Bioinformatics analyzed the expression and correlation of HJURP and FOXM1 in breast cancer tissues,and analyzed the enrichment pathway of HJURP.The expression of HJURP and FOXM1 in breast cancer cells and doxorubicin resistant cells were detected by qPCR.The binding relationship between FOXM1 and HJURP was verified by double Luciferase experiment and ChIP experiment.CCK-8 was used to detect cell viability and IC 50 values.Western blot was used to detect the expression of specific genes related to glycolytic metabolic pathways.Seahorse XP96 analyzed the extracellular acidification rate(ECAR)and oxygen consumption rate(OCR)of different treatment groups,and the assay kit detected the levels of pyruvate,lactate,and ATP in cells of each treatment group.Results HJURP was highly expressed in breast cancer and enriched in glycolysis pathway.Cell function experiments had found that HJURP could promote cell resistance to doxorubicin by regulating aerobic glycolysis.In addition,FOXM1 targets HJURP and was highly expressed in breast cancer,which was positively correlated with HJURP.In response experiment,HJURP overexpression could reverse the inhibition of FOXM1 knockdown on doxorubicin resistance in breast cancer cells.Conclusion The results of this study demonstrate that the transcription factor FOXM1 can activate the transcriptional regulation of HJURP aerobic glycolysis to promote doxorubicin resistance in breast cancer cells.