RANBP9调控TGF⁃β诱导结肠癌Colo320细胞凋亡的机制研究  

Ran⁃binding protein 9 targets transforming growth factor⁃β1 expression and induces cell apoptosis in colorectal cancer Colo320 cells

作　　者：班汝波 杨婕琳[2] 王晓媛 武雪亮[1] 路永刚[3] BAN Rubo;YANG Jielin;WANG Xiaoyuan;WU Xueliang;LU Yonggang(Department of General Surgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China;Department of Gastroenterology,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000;Laboratory,Hebei Provincial People’s Hospital,Shijiazhuang 050000)

机构地区：[1]普通外科,河北北方学院附属第一医院,河北张家口075000 [2]消化内科,河北北方学院附属第一医院,河北张家口075000 [3]实验室,河北省人民医院,河北石家庄050000

出　　处：《中国比较医学杂志》2024年第7期39-47,共9页Chinese Journal of Comparative Medicine

基　　金：河北省科技厅重点研发计划项目(2237784D);2020张家口市级科技计划自筹经费项目(2021050D)。

摘　　要：目的探讨Ran蛋白微管组织中心结合蛋白(RANBP9)靶向调控转化生长因子β1(transforming growth factor⁃β1,TGF⁃β1)的表达,及其对结直肠癌Colo320细胞凋亡的影响。方法分析TCGA数据库中625例结肠癌组织以及20例正常结肠组织中RANBP9的表达数据,KMPLOT分析RANBP9的表达与结肠癌患者生存期的关系。人类蛋白免疫组化数据库分析TGF⁃β1在正常结肠组织和结肠癌组织中的表达情况,使用UALCAN数据库分析TGF⁃β1的表达与结肠癌患者生存期的关系。双荧光素酶实验分析RANBP9和TGF⁃β1的关系。将pcDNA3.1⁃GFP⁃RANBP9和pcDNA3.1⁃GFP⁃RANBP9⁃NC分别转染至实验组和对照组细胞中,转染完成后细胞常规培养,另设立正常组细胞,不经过转染操作,常规培养;噻唑蓝(thiazolyl blue tetrazolium bromide,MTT)法检测各组细胞的生长活力,流式细胞仪检测各组细胞的凋亡率,JC⁃1染色检测各组细胞的线粒体膜电位,Western blot检测各组细胞中RANBP9和TGF⁃β1的表达。结果RANBP9在结肠癌组织中的表达明显降低,与RANBP9表达低的患者相比,RANBP9高表达的结肠癌患者具有较高的生存期曲线,TGF⁃β1在结肠癌组织的表达明显升高,与TGF⁃β1表达高的患者相比,TGF⁃β1低表达的患者具有较高的生存期曲线,差异均具有统计意义(P<0.05)。在结肠癌中,RANBP9能靶向调控TGF⁃β1的表达。与正常组细胞相比,实验组细胞的生长活力、线粒体的膜电位、TGF⁃β1的表达明显下调,细胞的凋亡率和RANBP9的表达明显升高,差异均具有统计意义(P<0.05)。结论RANBP9能靶向调控TGF⁃β1的表达,促使结肠癌细胞Colo320的生长活力和线粒体膜电位下降,诱导其凋亡。Objective To investigate the targeting of transforming growth factorβ1(TGF⁃β1)expression by Ran⁃binding protein 9(RANBP9)and its effect on colorectal cancer Colo320 cell apoptosis.Methods Gene expression levels of RANBP9 were analyzed in 625 colon cancer tissues and 20 normal colon tissues in The Cancer Genome Atlas database.The relationship between RANBP9 expression and survival time of patients with colon cancer was analyzed using KMPLOT.The expression of TGF⁃β1 in normal colon tissues and colon cancer tissues was analyzed using the human protein immunohistochemistry database and the relationship between TGF⁃β1 expression and the survival of patients with colon cancer was analyzed using the UALCAN database.The relationship between RANBP9 and TGF⁃β1 was analyzed by dual luciferase experiments.Colo320 cells were transfected with pcDNA3.1⁃GFP⁃RANBP9 and control pcDNA3.1⁃GFP⁃RANBP9⁃NC plasmids,respectively,and normal control cells were established without transfection.The cells were cultured and the growth viability of each group of cells was detected by the iazolyl blue tetrazolium bromide method,apoptosis was detected by flow cytometry,the mitochondrial membrane potential was detected by JC⁃1 staining,and RANBP9 and TGF⁃β1 protein expression were detected by Western blot.Results RANBP9 expression was significantly reduced in colon cancer tissues.Compared with patients with low RANBP9 expression,patients with high RANBP9 expression had a higher survival curve and significantly higher expression of TGF⁃β1 in colon cancer tissue.Compared with patients with high TGF⁃β1 expression,patients with low TGF⁃β1 expression had a significantly higher survival curve(P<0.05).RANBP9 targeted the expression of TGF⁃β1 in colon cancer.Compared with the normal group,cell growth,mitochondrial membrane potential,and TGF⁃β1 expression were all significantly down⁃regulated and the apoptosis rate and RANBP9 expression were significantly increased in the experiment group(P<0.05).Conclusionst

关 键 词：Ran蛋白微管组织中心结合蛋白 转化生长因子Β1 结直肠癌 细胞凋亡 

分 类 号：R-33[医药卫生]