siRNA沉默钙蛋白酶1对胆囊癌细胞增殖和迁移的影响  

Effects of siRNA Silencing of Calpain 1 on the Proliferation and Migration of Gallbladder Carcinoma Cells

作　　者：赵治艳 徐璟[1] 范璐璐[1] 笪洁[1] 刘萍萍[1] 彭万仁[1] 顾康生[1] Zhao Zhiyan;Xu Jing;Fan Lulu;Da Jie;Liu Pingping;Peng Wanren;Gu Kangsheng(Department of Oncology,the First Affiliated Hospital of Medical University of Anhui,Hefei 230000,China)

机构地区：[1]安徽医科大学第一附属医院肿瘤内科,合肥230000

出　　处：《中国药师》2019年第1期31-35,共5页China Pharmacist

摘　　要：目的:探究siRNA沉默钙蛋白酶1(Calpain1,CAPN1)对胆囊癌细胞GBC-SD增殖和迁移的影响及其潜在的分子机制。方法:将阴性siRNA、CAPN1-siRNA分别转染至细胞GBC-SD,以未转染的细胞为对照组,转染48 h后,采用荧光倒置显微镜检测转染效率,实时荧光定量PCR(qRT-PCR)检测GBC-SD细胞中CAPN1的表达情况,应用MTT实验检测GBC-SD细胞增殖能力,采用划痕实验检测GBC-SD细胞迁移能力,采用蛋白免疫印迹(Western blot)检测GBC-SD细胞中增殖细胞核抗原(PCNA)和CD44蛋白表达情况。结果:显微镜检测结果显示转染效率达到90%以上;与对照组、siRNA组相比,CAPN1-siRNA组细胞增殖率和迁移能力显著降低,CAPN1、PCNA、CD44蛋白表达显著降低(P <0. 05)。结论:siRNA沉默CAPN1具有一定的抑制胆囊癌细胞增殖和迁移作用,可为胆囊癌的治疗提供新的实验证据。Objective:To explore the effects of siRNA silencing of calpain 1(CAPN1)on the proliferation and migration of gallbladder carcinoma GBC-SD cells and the underlying molecular mechanisms.Methods:CAPN1-siRNA and siRNA were transfected into gallbladder cancer cell GBC-SD,and respectively named as CAPN1-siRNA group and siRNA group.The cells without transfection were served as the control.After 48 hours,a fluorescence microscope was used to detect the transfection efficiency,real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression of CAPN1 in GBC-SD cells,MTT and scratch assay were used to detect the proliferation and migration ability of GBC-SD cells,and Western blot was used to detect the expressions of CD44 and PCNA in GBC-SD cells.Results:The results showed that the transfection efficiency reached above 90%;the proliferation and migration rate in CAPN1-siRNA group was significantly lower than that in the control group(P<0.05).The expression levels of CAPN1,PCNA and CD44 were significantly lower than those in the control group(P<0.05).Conclusion:SiRNA silencing CAPN1 can inhibit the proliferation and migration of gallbladder cancer cells,which may provide new experimental evidence for the treatment of gallbladder cancer.

关 键 词：胆囊癌 钙蛋白酶1 细胞增殖 细胞迁移 增殖细胞核抗原 CD44 

分 类 号：R965.1[医药卫生—药理学]