干扰素/维甲酸联合应用诱导凋亡的相关基因19调控信号转导与转录激活因子3信号通路抑制肾癌细胞生长与增殖的机制研究  

Mechanism of GRIM-19 regulating signal transducer and activators of transcription 3 signaling pathway to inhibit the growth and proliferation of renal cell carcinoma

作　　者：崔林刚[1] 文建国[1,2] 何翔飞 袁鹏辉[1] 孟庆军 Cui Lingang;Wen Jiangguo;He Xiangfei;Yuan Penghui;Meng Qinjun(Department of Urology,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Urodynamics Center of Henan Province,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院泌尿外科,郑州450052 [2]河南省尿动力学中心,郑州450052

出　　处：《中华实验外科杂志》2024年第6期1156-1158,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨干扰素/维甲酸联合应用诱导凋亡的相关基因19(GRIM-19)抑制肾癌细胞生长与增殖的具体机制。方法取自于肾细胞癌患者肿瘤组织(30例,7例1期,8例2期,8例3期,7例4期),对照组取临近的正常组织(10例)。在肾癌细胞过表达GRIM-19后,应用RT-qPCR和蛋白质印迹法(Western blot)评估信号转导与转录激活因子3(STAT3)及其下游通路蛋白的表达水平变化,应用噻唑蓝(MTT)(药敏及细胞增殖实验)测定肾癌细胞增殖水平变化,应用流式细胞仪测定肾癌细胞的凋亡水平变化。结果与正常组织比较,GRIM-19在肾癌中表现为低表达,SATA3及其下游蛋白则表现为高表达。应用过表达GRIM-19腺病毒和空载腺病毒感染786-O细胞系,与空载腺病毒感染细胞组(1.62±0.07、1.47±0.08、1.23±0.50)比较,过表达GRIM-19腺病毒感染细胞组中GRIM-19(4.45±1.01)、B细胞淋巴瘤/白血病-2相关X蛋白(bax)(2.11±0.30)和半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-9(1.87±0.09)的表达水平显著提高。与空载腺病毒感染细胞组(1.68±0.70、0.91±0.30、1.38±0.20)比较,而STAT3(1.08±0.20)、B细胞淋巴瘤/白血病-XL(bcl-XL)(0.67±0.40)和细胞周期蛋白B1(Cyclin B1)(0.93±0.09)的表达水平明显降低。此外,干扰素(IFN)-β和ATRA可以抑制肾癌细胞的生长和诱导其凋亡,与IFN-β-GRIM-19组(15.30±2.60)、ATRA-GRIM-19(14.60±2.80)组比较,IFN-β-ATRA-GRIM-19组(21.70±3.10)显著地增加了(人肾细胞癌细胞系)786-O细胞的凋亡。结论肾细胞癌中GRIM-19的表达与STAT3诱导基因过度表达密切相关,GRIM-19通过调控STAT3信号通路抑制肾癌细胞生长与增殖。Objective To study the specific mechanism of GRIM-19 inhibiting the growth and proliferation of renal cell carcinoma(RCC).Methods The patients with renal cell carcinoma by nephrectomy in this urology department were selected from September 2021 to September 2022,and tumor tissues were obtained from patients with RCC(30 cases,7 patients with stage 1,8 patients with stage 2,8 patients with stage 3,and 7 patients with stage 4),and adjacent normal tissues were obtained from controls(10 cases).After overexpression of GRIM-19 in RCC cells,the expression levels of signal transducer and activators of transcription 3(STAT3)and its downstream pathway proteins were detected by RT-qPCR and Western blotting,and the proliferation level of RCC cells was determined by methyl thiazolyl tetrazolium(MTT)assay,the level of apoptosis was measured by flow cytometry.SPSS 20.0 was used for analysis.One-way ANOVA was used for sample mean comparison,and T-test was used for pairwise comparison.Results GRIM-19 was down-expressed in RCC cells as compared with normal tissues,while SATA3 and its downstream proteins were highly expressed.The expression levels of GRIM-19(4.45±1.01),B cell lymphoma/leukemia-2 associated X protein(bax)(2.11±0.30)and cysteinyl aspartate-specific protease(Caspase)-9(1.87±0.09)were significantly increased in the adenovirus infection cells overexpressing GRIM-19 as compared with those in the 786-O cells infected with empty adenovirus(1.62±0.07,1.47±0.08 and 1.23±0.50 respectively).The expression levels of STAT3,B cell lymphoma/leukemia-XL(bcl-XL)and Cyclin B1 in the adenovirus infection cells overexpressing GRIM-19 were significantly lower than those in the control group(1.68±0.70 vs.1.08±0.20;0.91±0.30 vs.0.67±0.40;1.38±0.20 vs.0.93±0.09).In addition,interferon(IFN)-βand ATRA could inhibit the growth and induce apoptosis of RCC cells.As compared with those of IFN-β-GRIM-19 group(15.30±2.60)and ATRA-GRIM-19 group(14.60±2.80),the apoptosis of 786-O cells in IFN-Β-ATRA-GRIM-19 group(21.70±3.10)increase

关 键 词：肾细胞癌 干扰素/维甲酸联合应用诱导凋亡的相关基因19 信号转导与转录激活因子3 增殖 凋亡 

分 类 号：R737.11[医药卫生—肿瘤]