摘要
目的研究肿瘤坏死因子受体相关因子相互作用蛋白(TRAIP)在人三阴性乳癌(TNBC)中的表达及其对细胞增殖和侵袭迁移的影响。方法应用免疫组化方法,检测75例TNBC癌及癌旁组织中TRAIP的表达,并分析其与病人临床病理特征的关系。采用Western blot技术检测人TNBC细胞系MDA-MB-468、MDA-MB-231及HCC1937中TRAIP的表达水平。构建TRAIP慢病毒干扰载体并检测干扰效率。应用3-(4,5-二甲基噻唑-2-基)-2,5-二苯四唑溴化铵(MTT)和Celigo细胞计数方法检测细胞增殖能力,Transwell侵袭实验和创面愈合法检测细胞的侵袭迁移能力。结果TNBC组织中TRAIP的表达水平明显高于癌旁组织(Z=-6.460,P<0.001);与原发灶相比,淋巴结转移灶中TRAIP呈高表达(Z=-3.448,P<0.001),并且这种高表达水平与肿瘤的复发密切相关(Z=-2.077,P<0.05)。Western blot检测显示,MDA-MB-231和HCC1937细胞株TRAIP蛋白表达均显著高于MDA-MB-468细胞(F=34.96,P<0.01);TRAIP基因敲除后,MDA-MB-231和HCC1937细胞中TRAIP的蛋白表达水平被明显抑制(t=15.66、19.39;P<0.01)。相较于对照组,TRAIP基因敲除组细胞的增殖能力明显减弱(F=71.63~218.07,P<0.01);Transwell侵袭及创面愈合实验表明,TRAIP基因敲除显著抑制细胞的侵袭迁移能力(t=7.39~32.16,P<0.01)。结论TRAIP在TNBC组织及淋巴结转移灶中高表达并与肿瘤的复发密切相关,敲除TRAIP基因可显著抑制细胞的增殖、侵袭迁移等生物学特性。
Objective To study the expression of tumor necrosis factor receptor-associated factor-interacting protein(TRAIP)in human triple-negative breast cancer(TNBC)and its role in cell proliferation,invasion,and migration.Methods We measured the expression of TRAIP in the cancer and paracancerous tissues from 75 cases of TNBC by immunohistochemistry,and analyzed its relationship with patients’clinicopathological features.Western blot was used to determine the expression of TRAIP in human TNBC cell lines(MDA-MB-468,MDA-MB-231,and HCC1937).We constructed a lentiviral vector targeting TRAIP,and measured its interference efficiency.Cell proliferation was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and Celigo cell counting assays.Cell invasion and migration was measured by Transwell and wound healing assays.Results The expression level of TRAIP was significantly higher in TNBC tissues than in paracancerous tissues(Z=-6.460,P<0.001),and significantly higher in lymph node metastases than in primary foci(Z=-3.448,P<0.001).A high TRAIP level was closely associated with tumor recurrence(Z=-2.077,P<0.05).The protein levels of TRAIP in the MDA-MB-231 and HCC1937 cell lines were significantly higher than that in the MDA-MB-468 cell line(F=34.96,P<0.01).After knocking out the TRAIP gene,the protein expression of TRAIP in the MDA-MB-231 and HCC1937 cell lines were significantly inhibited(t=15.66,19.39;P<0.01).Compared with the control group,the TRAIP knockout group showed significantly decreased cell proliferation(F=71.63-218.07,P<0.01)and significantly reduced cell invasion and migration(t=7.39-32.16,P<0.01).Conclusion TRAIP is highly expressed in TNBC tissues and lymph node metastases,and is closely related to tumor recurrence.Silencing the TRAIP gene can significantly inhibit cell proliferation,invasion,and migration.
作者
郑艳
王萍
王笑
刘俐彤
陈召旭
王成勤
ZHENG Yan;WANG Ping;WANG Xiao;LIU Litong;CHEN Zhaoxu;WANG Chengqin(Department of Pathology,School of Basic Medicine,Qingdao University,Qingdao 266071,China)
出处
《青岛大学学报(医学版)》
CAS
2023年第1期21-26,共6页
Journal of Qingdao University(Medical Sciences)
基金
国家自然科学基金资助项目(81672606)。
关键词
肿瘤坏死因子受体相关因子相互作用蛋白
三阴性乳腺癌
基因沉默
细胞增殖
肿瘤浸润
tumor necrosis factor receptor-associated factor interaction protein
triple negative breast neoplasms
gene silencing
cell proliferation
neoplasm invasiveness