泛素羧基末端水解酶37对胰腺癌SW1990细胞增殖的影响  

Effect of ubiquitin carboxyl-terminal hydrolase 37 on the proliferation of SW1990 pancreatic cancer cells

作　　者：闫帅 岳学良 杨森 谭飞龙 刘红山 YAN Shuai;YUE Xueliang;YANG Sen;TAN Feilong;LIU Hongshan(Department of Hepatobiliary Pancreatic Surgery,Zhengzhou University People's Hospital,Henan Provincial People's Hospital,Zhengzhou,Henan 450003,China)

机构地区：[1]郑州大学人民医院河南省人民医院肝胆胰腺外科,河南郑州450003

出　　处：《中华实用诊断与治疗杂志》2024年第5期433-437,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省医学科技攻关计划省部共建项目(SB201901079);河南省医学科技攻关计划联合共建项目(LHGJ20200011)。

摘　　要：目的 观察胰腺癌SW1990细胞中泛素羧基末端水解酶37(UCH37)对细胞分裂周期蛋白25A(CDC25A)的调控作用,探讨其对SW1990细胞增殖的影响。方法 取对数生长期正常人胰腺HPDE细胞及胰腺癌SW1990细胞,分别采用实时荧光定量PCR法、Western blot法检测UCH37 mRNA及蛋白相对表达量。取对数生长期SW1990细胞分为UCH37敲低组(转染sh-UCH37慢病毒)、UCH37空白组(转染UCH37-NC慢病毒)。转染48 h,分别采用实时荧光定量PCR法、Western blot法检测2组UCH37、CDC25A mRNA及蛋白相对表达量,采用细胞克隆形成实验检测2组细胞集落形成数;转染后培养24、48、72、96 h采用CCK-8法检测2组细胞增殖吸光度(optical density, OD)值。结果 (1)SW1990细胞UCH37 mRNA(3.338±0.137)及蛋白(0.956±0.043)相对表达量均高于HPDE细胞(0.968±0.040、0.056±0.003)(t=26.710,P<0.001;t=36.080,P<0.001)。(2)转染48 h, UCH37敲低组UCH37 mRNA(0.337±0.017)及蛋白(0.424±0.010)相对表达量均低于UCH37空白组(1.017±0.052、1.163±0.132)(t=21.810,P<0.001;t=9.160,P<0.001),细胞集落形成数[(27.0±7.9)个]少于UCH37空白组[(125.3±14.1)个](t=10.860,P<0.001)。UCH37敲低组转染48、72、96 h时OD值(0.433±0.016、0.563±0.004、0.897±0.046)均低于UCH37空白组(0.531±0.014、0.826±0.017、1.859±0.126)(t=7.420,P=0.002;t=24.750,P<0.001;t=12.300,P<0.001),转染24 h时OD值与UCH37空白组比较差异无统计学意义(P>0.05)。(3)转染48 h, UCH37敲低组CDC25A蛋白相对表达量(0.501±0.026)低于UCH37空白组(1.174±0.038)(t=23.090,P<0.001),CDC25A mRNA相对表达量与UCH37空白组比较差异无统计学意义(P>0.05)。结论 去泛素化酶UCH37可通过CDC25A调控胰腺癌细胞周期增殖,敲低UCH37表达可抑制胰腺癌细胞的增殖。Objective To observe the role of ubiquitin carboxyl-terminal hydrolase 37(UCH37)in regulating cell division cycle protein 25A(CDC25A)in SW1990 pancreatic cancer cells and to explore its effect on the proliferation of SW1990 cells.Methods The normal human pancreatic HPDE cells and pancreatic cancer SW1990 cells in logarithmic growth phase were harvested,and the relative expressions of UCH37 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot.The SW1990 cells were transfected with sh-UCH37 lentivirus(UCH37 knockdown group)and UCH37-NC lentivirus(UCH37 blank group)for 48 h.The relative expressions of UCH37 and CDC25A mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot.The cell colony formation assay was used to detect the number of colony formation.The CCK-8 method was used to detect the optical density(OD)values of cell proliferation after culture for 24,48,72 and 96 h in two groups.Results(1)The relative expressions of UCH37 mRNA and protein were higher in SW1990 cells(3.338±0.137,0.956±0.043)than those in HPDE cells(0.968±0.040,0.056±0.003)(t=26.710,P<0.001;t=36.080,P<0.001).(2)After culture for 48 h,the relative expressions of UCH37 mRNA and protein were lower in UCH37 knockdown group(0.337±0.017,0.424±0.010)than those inUCH37blank group(1.017±0.052,1.163±0.132)(t=21.810,P<0.001;t=9.160,P<0.001),and the number of colony formation was less in UCH37 knockdown group(27.0±7.9)than that in UCH37 blank group(125.3±14.1)(t=10.860,P<0.001).The OD values after culture for 48,72 and 96 h were lower in UCH37knockdown(0.433±0.016,0.563±0.004,0.897±0.046)than those in UCH37blank group(0.531±0.014,0.826±0.017,1.859±0.126)(t=7.420,P=0.002;t=24.750,P<0.001;t=12.300,P<0.001),andtherewasno significant difference in the OD value after culture for 24 h between two groups(P>0.05).(3)The relative expression of CDC25A protein was lower in UCH37 knockdown group(0.501±0.026)than that in UCH37 blank group(1.174±0.038)(t=23.090,P<0.001),while t

关 键 词：胰腺癌 泛素羧基末端水解酶37 细胞分裂周期蛋白25A 

分 类 号：R735.9[医药卫生—肿瘤]