微小RNA-205-5p调控轴抑制蛋白2基因表达对结肠癌细胞增殖凋亡的影响及作用机制的临床研究  

MicroRNA-205-5p participating in the proliferation and apoptosis of colon cancer cells by regulating the expression of axis inhibition protein 2 gene

作　　者：侯丽英[1] 张惠玲[1] 李霞[1] 侯文娅 董彦红 曹慧慧 Hou Liying;Zhang Huilin;Li Xia;Hou Wenya;Dong Yanhong;Cao Huihui(Department of Gastrointestinal Surgery,Shanxi Provincial Peoples Hospital,Taiyuan 030012,China)

机构地区：[1]山西省人民医院消化外科,太原030012

出　　处：《中华实验外科杂志》2021年第2期241-245,共5页Chinese Journal of Experimental Surgery

摘　　要：目的探讨微小RNA(microRNA,miR)-205-5p靶向调节轴抑制蛋白2(AXIN2)基因表达对结肠癌细胞增殖凋亡的影响及作用机制。方法收集2017年1月至2019年1月山西省人民医院手术切除结肠癌组织组织标本并购买结肠癌细胞系,分别以距肿瘤边缘5 cm的癌旁组织和人正常结肠黏膜上皮细胞系为对照。实时定量反转录聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)检测分别检测组织和细胞中miR-205-5p和AXIN2表达水平。荧光素酶报告实验验证miR-205-5p与AXIN2的靶向关系。细胞计数试剂盒(CCK-8)法和流式细胞术检测各组转染细胞增殖和凋亡变化,两组和多组间比较分别采用t检验和单因素方差分析。结果结肠癌组织及细胞系中miR-205-5p低表达(2.16±0.24比1.00±0.04,t=10.660,P<0.01),AXIN2高表达(mRNA,0.21±0.02比1.00±0.04,t=30.600,P<0.01;蛋白,0.64±0.03比1.44±0.06,t=26.670,P<0.05),差异均有统计学意义。AXIN2是miR-205-5p的靶基因。mimic组AXIN2蛋白表达量显著低于Blank组和NC组,差异有统计学意义(0.33±0.03,t=116.490,P<0.05);miR-205-5p抑制剂(inhibitor)组AXIN2蛋白表达量显著高于Blank组和NC组,差异有统计学意义(1.67±0.11,t=11.270,P<0.05)。miR-205-5p mimic组(增殖,48 h 0.82±0.09,t=5.375,P<0.05;72 h 1.46±0.13,t=3.333,P<0.05;凋亡,28.71±2.27,t=12.350,P<0.05)和AXIN2沉默表达(siRNA-AXIN2)组(增殖,48 h 1.11±0.11,t=2.814,P<0.05;72 h 1.68±0.06,t=4.959,P<0.05;凋亡,4.45±0.64,t=5.090,P<0.05)结肠癌细胞增殖率显著低于Blank组和NC组,细胞凋亡率显著高于Blank组和NC组,差异有统计学意义;而miR-205-5p inhibitor组(增殖,48 h 2.11±0.18,t=4.386,P<0.05;72 h 2.66±0.19,t=4.898,P<0.05;凋亡,4.71±0.26,t=5.155,P<0.05)和AXIN2过表达(OE-AXIN2)组(增殖,48 h 0.44±0.05,t=0.002,P<0.05;72 h 0.88±0.11,t=7.446,P<0.05;凋亡,25.65±2.12,t=10.900,P<0.05)结肠癌细胞增殖率显著高于Blank组和NC组,细胞凋亡率显著低于Blank组和NC组,差异有统计学意义。结论miR-2Objective To investigate the mechanism of microRNA(miRNA,miR)-205-5p on the proliferation and apoptosis of colon cancer cells by regulating the expression of axis inhibition protein 2(AXIN2)gene.Methods The specimens of colon cancer tissues were collected in our hospital from January 2017 to January 2019 and the colon cancer cell lines were purchased.The adjacent tissues 5 cm away from the edge of the tumor and the normal human colon epithelial cell line were used as controls.Fluorescent quantitative real-time polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the expression of miR-205-5p and AXIN2 in tissues and cells respectively.Luciferase Report experiment verified the targeting relationship between miR-205-5p and AXIN2.Cell counting kit-8(CCK-8)method and flow cytometry were used to detect the proliferation and apoptosis of transfected cells.Results There were low miR-205-5p expression(2.16±0.24 vs.1.00±0.04,t=10.660,P<0.01)and high AXIN2 expression(mRNA:0.21±0.02 vs.1.00±0.04,t=30.600,P<0.01;protein:0.64±0.03 vs.1.44±0.06,t=26.670,P<0.05)in colon cancer tissues and cell lines.AXIN2 was the target gene of miR-205-5p.Compared with Blank group and negative control(NC)group,there was significantly low AXIN2 expression in miR-205-5p mimic group,with statistical difference(0.33±0.03,t=116.490,P<0.01),while obviously high AXIN2 expression in miR-205-5p inhibitor group,with statistical difference(1.67±0.11,t=11.270,P<0.05).Compared with blank group and NC group,miR-205-5p mimic group(proliferation:48 h:0.82±0.09,t=5.375,P<0.05;72 h:1.46±0.13,t=3.333,P<0.05;apoptosis:28.71±2.27,t=12.350,P<0.05)and AXIN2 expression silencing(siRNA-AXIN2)group(proliferation:48 h:1.11±0.11,t=2.814,P<0.05;72 h:1.68±0.06,t=4.959,P<0.05;apoptosis:4.45±0.64,t=5.090,P<0.05)had inhibited proliferation of colon cancer cells and increased apoptosis rate,with statistical difference(all P<0.05).While compared with Blank group and NC group,miR-205-5p inhibitor group had promoted proliferation of colon cancer

关 键 词：微小RNA 结肠癌 增殖 脱噬作用 

分 类 号：R735.34[医药卫生—肿瘤]