EGFL6对人膀胱癌细胞体外及裸鼠皮下移植瘤增殖能力的影响  

Effects of EGFL6 on the proliferation of human bladder cancer cells in vitro and subcutaneously transplanted tumors in nude mice

作　　者：杨占锋[1] 王雷[1] 邱晓东[1] 李建华[1] 朱照伟[2] Yang Zhanfeng;Wang Lei;Qiu Xiaodong;Li Jianhua;Zhu Zhaowei(Department of Urology,Shangqiu First People’s Hospital,Shangqiu 476100,China;Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]商丘市第一人民医院泌尿外科,商丘476100 [2]郑州大学第一附属医院泌尿外科,郑州450052

出　　处：《中华内分泌外科杂志》2022年第6期677-680,共4页Chinese Journal of Endocrine Surgery

基　　金：国家自然科学基金(81702503)。

摘　　要：目的探究人表皮生长因子样结构域蛋白6(epidermal growth factor-like domain protein 6,EGFL6)基因在人膀胱癌细胞5637中的低表达对其体外和体内增殖能力的影响。方法将人膀胱癌细胞5637分为实验组和对照组,实验组细胞利用小干扰RNA(siRNA)靶向人EGFL6基因,对照组细胞转染Mock-siRNA;采用实时定量PCR法检测实验组和对照组细胞中EGFL6 mRNA的含量;利用CCK8检测各组细胞的增殖能力;将裸鼠皮下分别注射实验组和对照组人膀胱癌细胞5637,通过裸鼠皮下移植瘤实验检测细胞在体内的增殖能力;通过蛋白免疫印迹检测EGFL6、p-P13K、p-AKT的表达。结果实验组和对照组EGFL6表达量分别为(0.19±0.03)和(0.91±0.11),siRNA-EGFL6降低实验组人膀胱癌5637细胞中EGFL6的蛋白表达;CCK8结果发现实验组和对照组的吸光度分别为(1.558±0.152)、(2.287±0.182),实验组细胞增殖能力明显下降;裸鼠皮下移植瘤结果表明,实验组和对照组裸鼠瘤块体积分别为(1192.07±250.9)、(2280.5±600.1)μm^(3),质量分别为(0.66±0.31)、(1.52±0.48)g,实验组细胞4周之后形成的瘤块体积和质量均下降;蛋白免疫印迹实验结果发现,对照组和实验组p-P13K表达量分别为(0.79±0.14)和(0.33±0.09),p-AKT表达量分别为(0.93±0.13)和(0.28±0.06),证实实验组细胞较于对照组细胞p-P13K、p-AKT均表达下降。结论EGFL6低表达会通过P13K-AKT信号通路抑制人膀胱癌细胞5637的体内与体外增殖能力。Objective To explore the effect of low expression of human epidermal growth factor-like domain protein 6(EGFL6)gene in human bladder cancer cell 5637 on its proliferation ability in vitro and in vivo.Methods Human bladder cancer cells 5637 were divided into experimental group and control group.The experimental group cells targeted human EGFL6 gene with small interfering RNA(siRNA),and the control group cells were transfected with Mock-siRNA.The cells in the experimental group and the control group were detected by real-time quantitative PCR.The content of EGFL6 mRNA in the medium.CCK8 was used to detect the proliferation ability of cells.Nude mice were injected subcutaneously with human bladder cancer cells 5637 in the experimental and control groups respectively,and the proliferation ability of the cells in vivo was detected by subcutaneous transplantation tumor assay in nude mice.The expression of EGFL6,p-P13K,and p-AKT was detected by western blotting.Results The expression of EGFL6 was 0.19±0.03 and 0.91±0.11 in the experimental and control groups,respectively.siRNA-EGFL6 decreased the protein expression of EGFL6 in human bladder cancer 5637 cells in the experimental group.CCK8 results showed that the absorbance of the experimental group and the control group were 1.558±0.152 and 2.287±0.182,respectively.The results of subcutaneous tumor transplantation in nude mice showed that the volume of tumor in experimental group and control group was(1192.07±250.9)μm^(3) and(2280.5±600.1)μm^(3),respectively.The mass were(0.66±0.31)g and(1.52±0.48)g,respectively.The tumor volume and mass of the experimental group decreased after 4 weeks.The results of protein immunoblotting experiments revealed that the expression of p-P13K was 0.79±0.14 and 0.33±0.09 in the control and experimental groups,respectively,and the expression of p-AKT was 0.93±0.13 and 0.28±0.06,respectively,confirming that the expression of p-P13K and p-AKT were decreased in the experimental group of cells compared with the control group.Concl

关 键 词：膀胱癌 人表皮生长因子样结构域蛋白6 细胞增殖 

分 类 号：R737.14[医药卫生—肿瘤]