海南医科大学学报2025，Vol.31Issue(21)：1639-1651,13.DOI:10.13210/j.cnki.jhmu.20241212.002
敲低NSD2对三阴性乳腺癌细胞基因表达谱及紫杉醇化疗敏感性的影响研究
Effects of NSD2 knockdown on gene expression profile and chemosensitivity to paclitaxel in triple negative breast cancer cells
摘要
Abstract
Objective:To investigate the effects of NSD2 knockdown on gene expression profile and the chemosensitivity to paclitaxel(PTX)in triple negative breast cancer(TNBC)cells.Methods:Lentviral shRNA vector was used to mediate NSD2 gene knockdown in MDA-MB-231 cells.Gene expression microarrays were used to screen for differentially expressed genes and cluster analysis,GO and KEGG enrichment analysis were performed.Realtime PCR and Western blot were used to determine the amount of NSD2 mRNA and protein expression.Upon the treatment of PTX,the morphology of apoptotic cells was observed by Hoechst staining,and the apoptosis rate was assessed by Annexin V-FITC/PI staining.Western blot analysis was performed to detect the protein expression levels of differential genes associated with programmed cell death pathway.TIMER2.0,UALCAN,and Kaplan-Meier Plotter databases were used to analyze the expression of NSD2 and differential genes associated with pro-grammed cell death pathway in breast cancer,TNBC,and normal breast tissues,as well as their prognostic value in TNBC pa-tients who had received chemotherapy.Results:A total of 483 differentially expressed genes were screened by microarray,includ-ing 324 up-regulated genes and 159 down-regulated genes(P<0.01).GO and KEGG enrichment analysis showed that differential-ly expressed genes were significantly involved in programmed cell death signaling pathways such as apoptosis and necroptosis(P<0.05).Knockdown of NSD2 significantly increased the apoptosis rate of MDA-MB-231 cells upon PTX treatment,and the expres-sion levels of proteins related to programmed cell death,including TNFSF10,TNFRSF10B,FAS,CASP1,and Cleaved PARP1,were also significantly enhanced(P<0.05).Compared to normal breast tissues,the expression levels of NSD2 and PARP1 in breast cancer and TNBC were significantly up-regulated,while the expression levels of TNFSF10,FAS and CASP1 were significantly down-regulated(P<0.001).Among TNBC patients who had received chemotherapy,the relapse-free survival rate of patients with high NSD2 expression was lower than that of patients with low expression,whereas patients with high expres-sion of TNFRSF10B,FAS,or CASP1 had a better prognosis than those with low expression(P<0.05).Conclusion:Knock-down of NSD2 changes the gene expression profile of TNBC cells and increases the chemosensitivity to PTX by inducing pro-grammed death,providing a novel insight into the molecular targets of chemotherapy resistance in TNBC.关键词
三阴性乳腺癌/NSD2/基因表达谱/紫杉醇/化疗敏感性Key words
Triple-negative breast cancer/NSD2/Gene expression profile/Paclitaxel/Chemosensitivity分类
临床医学引用本文复制引用
陈艳,赵金辉,胡诗涵,王禹,潘云,高波..敲低NSD2对三阴性乳腺癌细胞基因表达谱及紫杉醇化疗敏感性的影响研究[J].海南医科大学学报,2025,31(21):1639-1651,13.基金项目
This study was supported by the National Natural Science Foundation of China(82160582) （82160582）
Yunnan Province Science and Technology Department Fundamental Research Project(202201AT070003) （202201AT070003）
Scientific Research Foundation of Dali(2021KBG036) （2021KBG036）
the First Affiliated Hospital of Dali University Discipline Building Key Project(DYFGG2022-01) （DYFGG2022-01）
the First Affiliated Hospital of Dali University Research Center Project(DFYXY009) 国家自然科学基金资助项目(82160582) （DFYXY009）
云南省科技厅基础研究计划面上项目(202201AT070003) （202201AT070003）
大理市科研基金资助项目(2021KBG036) （2021KBG036）
大理大学第一附属医院学科建设骨干项目(DFYGG2022-01) （DFYGG2022-01）
大理大学第一附属医院临床医学研究中心项目(DFYXY009) （DFYXY009）
