微小RNA-320a和钠氢交换调控因子1在肝细胞癌中的表达及机制  

Expression and mechanism of microRNA and Na+/H+exchanger regulatory factor 1 on hepatocellular carcinoma

作　　者：丁兢[1] 段斌炜[1] 邸亮[1] Ding Jing;Duan Binwei;Di Ling(Department of General Surgery,Beijing YouAn Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学附属北京佑安医院普外中心,北京100069

出　　处：《中国肝脏病杂志（电子版）》2020年第3期59-64,共6页Chinese Journal of Liver Diseases：Electronic Version

基　　金：首都临床特色应用研究(Z181100001718143)。

摘　　要：目的探讨微小RNA(microRNA,miRNA)-320a和钠氢交换调控因子1(Na+/H+exchanger regulatory factor 1,NHERF1)在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及机制。方法收集首都医科大学附属北京佑安医院2015年1月至2016年1月经手术治疗的HCC患者肝癌组织及癌旁组织。采用反转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)检测miR-320a和NHERFl在HCC组织、癌旁组织、HCC细胞株Bel-7402和正常肝细胞株HL-7702中的表达。将体外培养的Bel-7402细胞分为对照组、空白组和miR-320a转染组,空白组中仅加入2 ml全培养基;对照组加入2 ml空载体质粒;miR-320a组将稀释的miR-320a混合液加入到完全养基中,最终体积为2 ml;采用RT-PCR检测Bel-7402细胞中miR-320a、NHERF1及β-catenin的表达,采用流式细胞术检测Bel-7402细胞的凋亡,采用Transwell检测Bel-7402细胞迁移侵袭能力。结果miR-320a(0.51±0.01 vs 0.83±0.02)和NHERFl(0.78±0.02 vs 1.42±0.05)在肝癌组织中的表达均显著低于癌旁组织,差异有统计学意义(t值分别为-80.25、-68.05,P均<0.001)。miR-320a(0.75±0.03 vs 0.81±0.04)和NHERFl(0.79±0.05 vs 1.58±0.05)在Bel-7402细胞中的相对表达量均显著低于HL-7702,差异有统计学意义(t=-2.73,P=0.021,t=-27.60,P<0.001)。对照组、空白组和miR-320a转染组在Bel-7402细胞中miR-320a相对表达量分别为0.77±0.04、0.79±0.05和1.28±0.07,差异有统计学意义(H=11.66,P=0.003),miR-320a转染组显著高于对照组和空白组(H值分别为8.308、8.308,P值分别为0.004、0.004)。对照组、空白组和miR-320a转染组Bel-7402细胞中NHERFl相对表达量分别为0.82±0.04、0.70±0.04和1.46±0.06,差异有统计学意义(H=15.17,P=0.001),miR-320a转染组显著高于对照组和空白组(H值分别为8.337、8.308,P值分别为0.004、0.004)。空白组、对照组和miR-320a转染组Bel-7402细胞凋亡率分别为11.2%、11.4%、32.5%,差异有统计学意义(χ^2=9263.95,P<0.001)。其中miR-320Objective To investigate the expression and mechanism of microRNA(miRNA)-320a and Na+/H+exchanger regulatory factor 1(NHERF1)on hepatocellular carcinoma(HCC).Methods The liver cancer tissues and adjacent tissues of patients with HCC who were operated on in Beijing YouAn Hospital,Capital Medical University from January 2015 to January 2016 were collected.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of miR-320a and NHERF1 in HCC tissues,adjacent tissues,HCC cell line BEL-7402 and normal liver cell line HL-7702.BEL-7402 cells were divided into control group,blank group and miR-320a transfection group.The blank group was added with 2 ml medium;the control group was added with 2 ml plasmid;the miR-320a transfection group was added with diluted miR-320a mixture into the medium,and the final volume was 2 ml.RT-PCR was used to detect the expression of miR-320a,NHERF1 andβ-catenin in Bel-7402 cells,flow cytometry was used to detect the apoptosis of Bel-7402 cells,and Transwell was used to detect the migration ability of Bel-7402 cells.Results The expression of miR-320a(0.51±0.01 vs 0.83±0.02)and NHERFl(0.78±0.02 vs 1.42±0.05)in liver cancer tissues were significantly lower than those in adjacent tissues,the differences were statistically significant(t=-80.25,-68.05;P<0.001).The expression of miR-320a(0.75±0.03 vs 0.81±0.04)and NHERFl(0.79±0.05 vs 1.58±0.05)in Bel-7402 cells were significantly lower than those in HL-7702 cells,the differences were statistically significant(t=-2.73,P=0.021;t=-27.60,P<0.001).The expression of miR-320a in Bel-7402 cells in control group,blank group and miR-320a transfection group were 0.77±0.04,0.79±0.05 and 1.28±0.07,the difference was statistically significant(H=11.66,P=0.003).The expression of miR-320a in Bel-7402 cells in miR-320a transfection group was significantly higher than those in control group and blank group(H=8.308,8.308;P=0.004,0.004).The expression of NHERFl in Bel-7402 cells in control group,blank group and miR-320a tran

关 键 词：肝细胞癌 微小RNA-320a 钠氢交换调控因子1 表达 WNT信号转导通路 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]