细胞迁移诱导蛋白对胆囊癌GBC-SD细胞生物学功能的影响及其机制研究  

Effect of cell migration-inducing hyaluronidase 1 on biological function of gallbladder cancer GBC-SD cells and its mechanism

作　　者：齐蕾[1] 陈文辉 叶健文[2] Qi Lei;Chen Wenhui;Ye Jianwen(Department of Pharmacology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052 [2]郑州大学第一附属医院肝胆胰外科,郑州450052

出　　处：《中华肝胆外科杂志》2022年第10期772-777,共6页Chinese Journal of Hepatobiliary Surgery

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20190028);河南省高等学校重点科研项目(19B320039)。

摘　　要：目的:探讨细胞迁移诱导蛋白(CEMIP)对胆囊癌细胞系GBC-SD生物学功能的影响及其可能机制。方法:检测CEMIP在胆管上皮细胞HIBEC及胆囊癌细胞系NOZ、GBC-SD中的表达。取对数生长期的人胆囊癌细胞系GBC-SD作为空白对照组;转染无意义的小干扰RNA作为阴性对照组;将siCEMIP-1或siCEMIP-2转染至GBC-SD细胞中,分别作为siCEMIP-1组和siCEMIP-2组。蛋白质印迹法检测GBC-SD细胞中CEMIP、免疫球蛋白重链结合蛋白(Bip)、钙网蛋白(CRT)的表达量,CCK-8法检测GBC-SD细胞相对存活率。分别采用划痕实验、流式细胞术检测细胞划痕修复率及细胞凋亡率。Transwell小室实验检测细胞迁移、侵袭能力。选取12只5周龄BALB/c-nude小鼠分为对照组和实验组,每组6只,两组小鼠分别于右侧腋下(前肢)皮下注射GBC-SD细胞和沉默CEMIP的GBC-SD细胞,33 d后比较移植瘤体积和重量。结果:与HIBEC细胞相比,GBC-SD[(0.750±0.034)比(0.120±0.002)]和NOZ[(0.690±0.013)比(0.120±0.002)]细胞中CEMIP蛋白相对表达量均增高,差异具有统计学意义(P<0.05)。与空白对照组及阴性对照组比较,siCEMIP-1组及siCEMIP-2组CEMIP、Bip及CRT的表达量、细胞相对存活率、细胞划痕修复率、细胞迁移及侵袭能力均下降,而细胞凋亡率增加,差异有统计意义(P<0.05)。与对照组相比,实验组小鼠的瘤体体积[(543.6±114.7)比(801.5±256.3)mm^(3)]和瘤体重量[(0.453±0.093)比(0.728±0.278)g]均下降,差异有统计学意义(P<0.05)。结论:CEMIP在胆囊癌细胞系GBC-SD及NOZ中高表达,沉默CEMIP基因表达可抑制胆囊癌细胞系GBC-SD增殖、划痕修复能力、迁移及侵袭能力,促进细胞凋亡,这可能与CEMIP抑制内质网分子伴侣Bip及CRT表达相关。Objective To investigate the effect of cell migration-inducing hyaluronidase 1(CEMIP)on biological function of gallbladder cancer GBC-SD cells and its possible mechanism.Methods The expression of CEMIP in biliary epithelial cell line HIBEC and gallbladder cancer cell line NOZ and GBC-SD was detected by Western blot.GBC-SD cells in logarithmic growth phase were divided into blank control group,negative control group(transfection with nonsense siRNA),siCEMIP-1 group(transfection with siCEMIP-1)and siCEMIP-2 group(transfection with siCEMIP-2).The expression of CEMIP and binding immunoglobulin protein(Bip)and calreticulin(CRT)in GBC-SD cells was detected by Western blot after culturing for 48h in blank control group,negative control group,siCEMIP-1 and siCEMIP-2 group.The relative survival rate was determined by CCK-8 assay.The wound healing rate and apoptotic rate was detected by wound healing assay and flow cytometry.The migration and invasion abilities were evaluated by Transwell chamber assay.Twelve 5-week-old BALB/c-nude mice were selected and divided into control group and experimental group(6 mice in each group).GBC-SD cells and GBC-SD cells with silenced CEMIP were subcutaneously injected into the right armpit(forelimb)of the two groups of mice,respectively.The volume and weight of transplanted tumor were compared 33 days later.Results Compared with HIBEC cells,the relative protein level of CEMIP in gallbladder cancer GBC-SD cells[(0.750±0.034)vs.(0.120±0.002)]and NOZ cells[(0.690±0.013)vs.(0.120±0.002)]was significantly higher(P<0.05).Compared with blank control group and negative control group,the relative protein level of CEMIP,Bip and CRT in siCEMIP-1 group and siCEMIP-2 group was significantly lower(P<0.05).Compared with blank control group and negative control group,the relative survival rate and wound healing rate and number of migration cells and invading cells in siCEMIP-1 group and siCEMIP-2 group were also significantly lower(P<0.05).While the apoptotic rate in siCEMIP-1 group and siCEMIP-2 gro

关 键 词：癌 胆囊 细胞迁移诱导蛋白 生物学功能 内质网应激 

分 类 号：R735.8[医药卫生—肿瘤]