NGAL基因沉默对人卵巢癌细胞株A2780凋亡的影响及机制  

Effect and mechanism of NGAL gene silencing on the apoptosis of human ovarian cancer cell line A2780

作　　者：刘荣 李忠梅[2] 熊海军[2] 曾瑞霞 LIU Rong;LI Zhong-mei;XIONG Hai-jun;ZENG Rui-xia(Department of Anatomy,Basic Medical College of Jinzhou Medical University,Jinzhou 121000,China;Laboratory Department of Fuming General Hospital of Liaoning Health Industry Group,Seventh Clinical College of China Medical University,Fushun 113008,China)

机构地区：[1]锦州医科大学基础医学院解剖学教研室,辽宁锦州121000 [2]辽宁省健康产业集团抚矿总医院,中国医科大学第七临床学院检验科,辽宁抚顺113008

出　　处：《解剖科学进展》2020年第3期328-331,335,共5页Progress of Anatomical Sciences

基　　金：辽宁省教育厅自然科学基金(L2015313)。

摘　　要：目的探讨人中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)基因沉默对人卵巢癌细胞株A2780凋亡的影响及机制。方法构建针对NGAL基因的siRNA真核表达载体,转染入A2780细胞中,实验分为3组:空白对照组(Control)、转染NGAL-siRNA沉默组(NGAL-siRNA)和转染阴性对照组(NC-NGAL)。免疫荧光检测NGAL在A2780细胞及IOSE80正常卵巢细胞的表达情况。通过RNA技术沉默NGAL后,运用qRT-PCR及Western blot检测沉默NGAL基因效率。CCK-8法检测细胞活力,TUNEL染色及流式细胞仪检测细胞凋亡率,Western blot检测NF-κB、Caspase-3蛋白表达。结果NGAL在A2780细胞呈高水平表达。NGAL-siRNA组NGAL基因及蛋白表达都受到明显的抑制(P<0.01)。与Control组相比,NGAL-siRNA组细胞凋亡率及Caspase-3蛋白表达明显增加,细胞活力及NF-κB蛋白表达明显下降(P<0.01),而NC-NGAL组各项指标与Control组相比差异无统计学意义(P>0.05)。结论沉默人卵巢癌细胞株A2780中NGAL基因表达,抑制细胞生长并促进细胞凋亡,与上调Caspase-3蛋白表达和下调NF-κB蛋白表达相关。Objective To investigate the effect and mechanism of neutrophil gelatinase-associated lipocalin(NGAL)gene silencing on the apoptosis of human ovarian cancer cell line A2780.Methods The eukaryotic expression vector of siRNA targeting NGAL gene was constructed and transfected into A2780 cells.The experiment was divided into 3 groups:blank control group(Control),NGAL-siRNA transfection group(NGAL-siRNA)and Transfection negative control group(NC-NGAL).The expressions of NGAL in A2780 cells and IOSE80 cells were detected by immunofluorescence.After NGAL gene silencing by siRNA,the efficiency of silencing NGAL gene was detected by qRT-PCR and Western blot.The cell viability was detected by CCK-8,the apoptosis rate was detected by TUNEL staining and flow cytometry,and the expression of NF-κB and Caspase-3 was detected by Western blot.Results NGAL was highly expressed in A2780 cells.The expression level of NGAL gene and protein was significantly inhibited in NGAL-siRNA group(P<0.01).Compared with control group,the apoptotic rate and expression level of Caspase-3 protein were significantly increased,cell viability and the expression level of NF-κB were significantly decreased(P<0.01)in NGAL-siRNA group,but with no significant difference between NC-NGAL group and control group(P>0.05).Conclusion Silencing NGAL gene in human ovarian cancer cell line A2780 inhibited cell growth and promoted cell apoptosis,which is related to upregulation of Caspase-3 expression and downregulation of NF-κB expression.

关 键 词：卵巢癌 基因沉默 NGAL基因 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]