干扰素诱导基因IFIT3对膀胱癌细胞生物学功能及化疗耐药性的影响  

The Mechanism of Interferon-inducible Gene IFIT3 in Chemotherapy Resistance of Bladder Cancer

作　　者：徐志成 秦卫军 杨力军 韩东辉 田春娟 XU Zhi-cheng;QIN Wei-jun;YANG Li-jun;HAN Dong-hui;TIAN Chun-juan(Department of Urology,The First Affiliated Hospital,Airforce Medical University of PLA,Xi'an,Shaanxi,710032,China)

机构地区：[1]中国人民解放军空军军医大学第一附属医院泌尿外科,陕西西安710032

出　　处：《现代生物医学进展》2021年第21期4011-4018,共8页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81772734)。

摘　　要：目的:探讨干扰素诱导基因IFIT3在膀胱癌中的表达及对膀胱癌细胞生物学功能和化疗耐药性的影响。方法:本研究通过q RT-PCR或Western blot检测了30例膀胱移行细胞癌患者的肿瘤组织和配对正常癌旁组织标本以及人膀胱癌细胞系T24及人膀胱上皮永生化细胞SV-HUC-1中IFIT3的表达水平。通过浓度递增法构建顺铂(DDP)耐药T24细胞(T24/DDP),然后对细胞转染靶向IFIT3的si RNA(si-IFIT3)及阴性对照(si-NC)。将细胞分为4组,分别为T24-si-NC组、T24-si-IFIT3组、T24/DDP-si-NC组、T24/DDP-si-IFIT3组,通过MTT法检测细胞增殖,使用Annexin V-FITC/PI凋亡检测试剂盒检测细胞凋亡,使用Transwell法检测细胞侵袭能力。通过qRT-PCR或Western blot检测HSP90α(HSP90AA1)、MMP2、MMP9、Cleaved-caspase-3、Bcl-2和Bax的表达水平。结果:与配对癌旁组织和SV-HUC-1相比,膀胱癌组织和T24细胞中IFIT3的m RNA和蛋白表达水平显著升高(P<0.001)。与正常T24细胞相比,T24/DDP细胞中的IFIT3 m RNA和蛋白表达水平显著升高(P<0.001)。与T24/DDP-si-NC组相比,T24/DDP-si-IFIT3组的IC_(50)值和侵袭细胞数显著降低,而细胞凋亡率显著升高(P<0.05)。与T24/DDP-si-NC组相比,T24/DDP-si-IFIT3组的Bcl-2、MMP2和MMP9蛋白相对表达量显著降低,而Bax和Cleaved-caspase-3蛋白相对表达量显著升高(P<0.05)。String数据库显示,IFIT3与HSP90AA1基因存在互作关系。与T24/DDP-si-NC组相比,T24/DDP-si-IFIT3组的HSP90α(HSP90AA1)表达水平显著降低(P<0.05)。结论:下调IFIT3的表达可抑制膀胱癌细胞的增殖和侵袭,并促进细胞凋亡,下调IFIT3可部分通过抑制HSP90α(HSP90AA1)的表达来降低膀胱癌细胞对顺铂的耐药性。Objective:To investigate the expression of interferon-induced gene IFIT3 in bladder cancer and its influence on the biological function and chemotherapy resistance of bladder cancer cells.Methods:In this study,IFIT3 expression in 30 bladder transitional cell carcinoma patients’tumor tissues and paired normal adjacent tissue specimens as well as human bladder cancer cell line T24 and human bladder epithelial immortalized cells SV-HUC-1 were detected by qRT-PCR or Western blot.The cisplatin(DDP)resistant T24 cells(T24/DDP)were constructed by the concentration increasing method,and then the cells were transfected with si RNA targeting IFIT3(si-IFIT3)and a negative control(si-NC).The cells were divided into 4 groups,namely T24-si-NC group,T24-si-IFIT3 group,T24/DDP-si-NC group,T24/DDP-si-IFIT3 group.MTT method was used to detect cell proliferation,Annexin V-FITC/PI apoptosis detection kit was used to detect cell apoptosis,and Transwell method was used to detect cell invasion ability.The expressions of HSP90α(HSP90 AA1),MMP2,MMP9,Cleaved-caspase-3,Bcl-2 and Bax were detected by qRT-PCR or Western blot.Results:Compared with paired adjacent tissues and SV-HUC-1,the expression levels of IFIT3 m RNA or protein in bladder cancer tissues and T24 cells were significantly increased(P<0.001).Compared with normal T24 cells,IFIT3 m RNA and protein expression levels in T24/DDP cells were significantly increased(P<0.001).Compared with the T24/DDP-si-NC group,the IC_(50) value and the number of invaded cells in the T24/DDP-si-IFIT3 group were significantly reduced,while the apoptosis rate was significantly increased(P<0.05).Compared with the T24/DDP-si-NC group,the relative expression of Bcl-2,MMP2 and MMP9 protein in the T24/DDP-si-IFIT3 group was significantly reduced,while the relative expression of Bax and Cleaved-caspase-3 protein was significantly increased(P<0.05).The String database showed that there was an interaction between IFIT3 and HSP90 AA1 genes.Compared with the T24/DDP-si-NC group,the expression level of HSP90α

关 键 词：干扰素诱导基因 IFIT3 膀胱癌 化疗耐药性 热休克蛋白90 

分 类 号：R-33[医药卫生] R737.14