摘要
目的:寻找糖酵解相关的他莫昔芬耐药(tamoxifen resistance,TamR)基因标志物,为乳腺癌(breast cancer,BC)的耐药机制提供理论依据。方法:从基因表达数据库GEO下载GSE67916,差异分析后与糖酵解相关基因集取交集,通过GSEA富集分析找到与他莫昔芬耐药相关的糖酵解基因,后续在外部独立数据集GSE125738和GSE9893对耐药基因进行验证。此外,在构建的他莫昔芬耐药细胞株中,对耐药基因使用qRT-PCR和Western blot技术在mRNA和蛋白层面进行验证,并通过CCK-8实验进一步验证耐药基因。结果:差异分析得到730个差异表达基因(differentially expressed genes,DEGs),与糖酵解相关基因集取交集得到19个DEGs,对这19个DEGs进行GSEA富集,我们发现PYGL与他莫昔芬耐药信号通路和内分泌治疗抵抗信号通路相关。在数据集GSE125738和GSE9893验证PYGL在他莫昔芬耐药组中表达上调。在乳腺癌他莫昔芬耐药细胞株中,PYGL的mRNA水平和蛋白水平均表达上调。此外,CCK-8实验表明,降低PYGL的表达可增强MCF7-TamR对他莫昔芬的反应性。结论:糖酵解基因PYGL是潜在的乳腺癌他莫昔芬耐药基因,可能为乳腺癌治疗提供新的认识和靶向策略。
Objective:To identify glycolysis-related gene markers associated with tamoxifen resistance(TamR)in breast cancer(BC)and provide a theoretical basis for understanding the mechanism of drug resistance.Methods:We downloaded the dataset GSE67916 from the Gene Expression Omnibus(GEO)database and performed differential analysis.The differentially expressed genes(DEGs)were intersected with the glycolysis-related gene set,and Gene Set Enrichment Analysis(GSEA)was conducted to identify glycolysis-related genes associated with tamoxifen resistance.The identified resistant genes were further validated in two external independent dataset,GSE125738 and GSE9893.In addition,qRT-PCR and Western blot techniques were used to validate the mRNA and protein expression of the resistant genes in the constructed tamoxifen-resistant cell line.CCK-8 experiments were conducted to further validate the role of resistant genes.Results:A total of 730 DEGs were obtained from the differential analysis,and 19 DEGs were identified by intersecting with the glycolysis-related gene set.GSEA enrichment analysis of these 19 DEGs revealed that PYGL were associated with tamoxifen resistance signaling pathway and endocrine therapy resistance signaling pathway.The upregulation of PYGL in the tamoxifen-resistant group was validated in the datasets GSE125738 and GSE9893.In the tamoxifen-resistant breast cancer cell line,both the mRNA and protein levels of PYGL were upregulated.Furthermore,CCK-8 experiments demonstrated that downregulation of PYGL expression enhanced the reactivity of MCF7-TamR to tamoxifen.Conclusion:Glycolytic gene PYGL is a potential tamoxifen resistance gene in breast cancer,which may provide new understanding and targeting strategies for breast cancer therapy.
作者
夏佳琳
赵卫卫
李静
XIA Jialin;ZHAO Weiwei;LI Jing(School of Laboratory Medicine and Biotechnology,Southern Medical University,Guangdong Guangzhou 510515,China;Affiliated Sixth People's Hospital South Campus,Shanghai Jiao Tong University,Shanghai 201400,China)
出处
《现代肿瘤医学》
CAS
2024年第20期3857-3863,共7页
Journal of Modern Oncology
基金
上海市奉贤区科委科技发展基金项目(编号:20211609)。
