CAPN1对胆囊癌细胞增殖、侵袭和迁移的影响  

Influences of CAPN1 on proliferation,invasion and migration of gallbladder carcinoma cell

作　　者：牛志鹏[1] 位亚秋 冯新献[1] 杨会锋[1] NIU Zhi-peng;WEI Ya-qiu;FENG Xin-xian;YANG Hui-feng(The First People＇s Hospital of Zhengzhou,Zhengzhou,Henan 450003,Chin)

机构地区：[1]郑州市第一人民医院,河南郑州450003

出　　处：《热带医学杂志》2018年第7期897-901,912,F0003,共7页Journal of Tropical Medicine

摘　　要：目的检测钙蛋白酶1(CAPN1)在胆囊癌(GBC)组织中的表达水平,探究其对人GBC细胞系(GBC-SD)增殖、侵袭及转移的影响。方法采用免疫组化法检测GBC组织、癌旁组织、正常胆囊组织中CAPN1蛋白表达水平,分析其与GBC患者临床病理参数的关系。体外培养GBC-SD细胞和正常胆囊上皮细胞,将GBC-SD细胞分为CAPN1-si RNA组、阴性对照组和空白对照组,病毒感染72 h后,采用实时荧光定量PCR(q RT-PCR)和蛋白免疫印迹(WB)技术检测细胞中CAPN1 m RNA及蛋白表达水平,CKK-8法检测细胞增殖情况,Transwell实验检测细胞迁移、侵袭能力。结果 GBC组织、癌旁组织、正常组织中CAPN1阳性表达率比较,差异有统计学意义(P<0.05)。不同TNM分期、病理分化和有无远处转移的GBC患者CAPN1阳性表达率比较,差异有统计学意义(P<0.05)。GBCSD细胞中CAPN1 m RNA及蛋白水平显著高于正常胆囊上皮细胞(P<0.05),感染CAPN1-si RNA后,GBC-SD细胞中CAPN1 m RNA及蛋白表达显著下调(P<0.05)。与空白及阴性对照组比较,si RNA组GBC-SD细胞的增殖、迁移、侵袭能力均显著下降(P<0.05)。结论 CAPN1在GBC组织中高表达,CAPN1沉默可抑制GBC细胞的增殖、迁移及侵袭,可能作为潜在生物靶标应用于GBC基因治疗。Objective To detect the expression level of calpain 1（CAPN1）in gallbladder carcinoma（GBC）tissue,and explore its influences on GBC-SD cell proliferation,invasion and migration of GBC cell line.Methods The expression levels of CAPN1 protein in GBC tissue,paracancerous tissue and normal gallbladder tissue were detected by immunohistochemistry,and analyzed its relationship with the clinicopathological parameters of GBC patients.GBC-SD cells and the normal gallbladder epithelial cells were cultured in vitro.GBC-SD cells were divided into CAPN1-Si RNA group,negative control group and blank control group;after 72 h of the virus infection,the expressions of CAPN1 m RNA and protein were detected by real time fluorescence quantitative PCR（q RT-PCR） and protein immunoblotting（WB）,respectively.CKK-8 assay was used to detect cell proliferation;transwell test was used to detect cell migration and invasion abilities.Results The differences in the positive expression rates of CAPN1 between the GBC tissue,paracancerous tissue and normal tissue were statistically significant（P〈0.05）.The differences in the positive expression rates of CAPN1 between the GBC patients with different TNM staging,pathological differentiation and distant metastasis were statistically significant（P〈0.05）.The levels of CAPN1 m RNA and protein in GBC-SD cells were significantly higher than those in normal gallbladder epithelial cells（P〈0.05）;after transfection of CAPN1-si RNA,the expressions of CAPN1 m RNA and protein in GBC-SD cells decreased significantly（P〈0.05）.Compared with the blank group and negative control group,the abilities of proliferation,migration and invasion of GBC-SD cells in Si RNA group were significantly decreased（P〈0.05）.Conclusion CAPN1 was highly expressed in GBC tissues,and CAPN1 silence could inhibit the proliferation,migration and invasion of GBC cells,and it might be used as a potential biological target for GBC gene therapy.

关 键 词：钙蛋白酶1 胆囊癌 细胞增殖 

分 类 号：R735.8[医药卫生—肿瘤]