c-Myc调控葡萄糖转运蛋白4激活酪氨酸激酶/信号转导与转录激活因子促进三阴性乳腺癌机制研究  

c-Myc regulates glucose transporters 4 to activate the janus kinase/signal transducer and activators of transcription to promote triple negative breast cancer

作　　者：张莹[1] 范静婧 吴中语 马伟强 马斌林[1] Zhang Ying;Fan Jingjing;Wu Zhongyu;Ma Weiqiang;Ma Binlin(Xinjiang Medical University Affiliated Tumor Hospital,the Clinical Medical Research Center of Breast and Thyroid Tumor in Xinjiang,Urumqi 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院,新疆乳腺甲状腺肿瘤临床医学研究中心,乌鲁木齐830011

出　　处：《中华实验外科杂志》2024年第4期674-678,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82260549)。

摘　　要：目的探讨c-Myc调控葡萄糖转运蛋白4(GLUT4)激活酪氨酸激酶(JAK)/信号转导与转录激活因子(STAT)信号通路促进三阴性乳腺癌(TNBC)进展机制。方法通过生信分析GLUT4、c-Myc关联性及GLUT4与乳腺癌临床病理特征间关系。收集三阴性乳腺癌患者临床样本,免疫组织化学检测GLUT4和c-Myc表达水平。构建敲除c-Myc的MDA-MB-231细胞株,通过实时定量反转录聚合酶链反应(RT-qPCR)及蛋白质印迹法(Western blot)检测GLUT4表达水平。构建敲除GLUT4和同时敲除GLUT4及c-Myc的MDA-MB-231细胞株,细胞计数试剂盒(CCK-8)检测增殖,流式细胞仪检测凋亡,Transwell检测迁移和侵袭。Western blot进一步分析c-Myc调控GLUT4对JAK/STAT的影响。正态分布计量资料比较采用独立样本t检验,多组资料组间比较采用方差分析。结果生信分析结果显示,在乳腺癌中GLUT4明显下调(P<0.01)且与乳腺癌TNM分期、病理分级、PAM50分型等相关。免疫组织化学评分结果显示,c-Myc在TNBC癌组织中表达显著高于癌旁组织(7.233比4.567,t=4.551,P<0.01),而GLUT4显著低表达(4.800比7.933,t=5.702,P<0.01)。在MDA-MB-231细胞中敲除c-Myc,RT-qPCR结果显示,c-Myc敲除组GLUT4表达量高于阴性对照组(2.486±0.273比1.070±0.140,F=55.005,P<0.01),Western blot结果显示,c-Myc敲除组GLUT4表达量高于阴性对照组(0.691±0.041比0.431±0.055,F=29.027,P<0.01)。构建GLUT4低表达的MDA-MB-231细胞株,RT-qPCR结果显示,GLUT4沉默组表达量低于阴性对照组,差异有统计学意义(0.507±0.076/0.137±0.025/0.317±0.085比1.150±0.200,F=45.016,P<0.01)。CCK-8结果显示,组细胞存活率高于阴性对照组(128.614±13.780比95.875±9.130,F=26.927,P<0.01),继续敲除c-Myc后细胞存活率低于阴性对照组(70.977±5.906比95.875±9.130,F=26.927,P<0.01)。流式细胞术结果显示,同时敲除GLUT4及c-Myc后细胞凋亡高于阴性对照组(12.170±1.283比3.263±0.691,F=102.946,P<0.01)。Transwell结果显示,敲除GLUT4后细胞迁�Objective To analyze the mechanism of c-Myc regulating glucose transporters 4(GLUT4)to activate janus kinase(JAK)/signal transducer and activators of transcription(STAT)to promote the progression of triple negative breast cancer(TNBC),and to explore new therapeutic targets.Methods The relationship between GLUT4 and c-Myc and the clinicopathological features of breast cancer were analyzed by bioinformatics.Clinical samples of TNBC patients were collected,and the expression levels of GLUT4 and c-Myc were detected by immunohistochemistry.MDA-MB-231 cell line with c-Myc knockout was constructed,and the expression of GLUT4 was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and Western blotting.Cell counting kit-8(CCK-8)assay was used to detect proliferation,flow cytometry was used to detect apoptosis,and Transwell assay was used to detect migration and invasion.Western blotting was used to further analyze the effect of c-Myc regulating GLUT4 on JAK/STAT.The measurement data of the two groups of normal distribution data were compared using the independent sample t test,and the data between numerous groups were compared using analysis of variance.Results Bioinformatics analysis showed that GLUT4 was significantly down-regulated in breast cancer(P<0.01)and correlated with TNM stage,pathological grade and PAM50 subtype.According to the immunohistochemical score,the expression levels of c-Myc in TNBC cancer tissues were significantly higher than paracancerous tissues(7.233 vs.4.567,t=4.551,P<0.01),while GLUT4 expression was significantly lower(4.800 vs.7.933,t=5.702,P<0.01).When c-Myc was knocked out in MDA-MB-231 cells,RT-qPCR results showed that the expression of GLUT4 in c-Myc knockout group was higher than that in negative control group(2.486±0.273 vs.1.070±0.140,F=55.005,P<0.01).Western blotting results showed that the expression of GLUT4 in c-Myc knockout group was higher than that in negative control group(0.691±0.041 vs.0.431±0.055,F=29.027,P<0.01).MDA-MB-231 cell lin

关 键 词：三阴性乳腺癌 C-MYC 葡萄糖转运蛋白4 酪氨酸激酶/信号转导与转录激活因子信号通路 

分 类 号：R737.9[医药卫生—肿瘤]