SATB2与miR-31、182对结直肠癌的靶向调控  

Detection of target regulation of SATB2, miR-31 and 182 in colorectal cancer

作　　者：杨洲[1] 刚海菊 覃先蓬[1] 贾贵清[1] 周晓刚[1] 赵高平[1] 高本领 YANG Zhou;GANG Hai-Ju;QIN Xian-Peng(Gastrointestinal Surgery, Sichuan Provincial People's Hospital, Chengdu 610000, Sichuan, China)

机构地区：[1]四川省人民医院胃肠外科,四川成都610000 [2]成都市职业技术学院医护分院护理教研室 [3]广元市苍溪县中医院

出　　处：《中国老年学杂志》2019年第7期1652-1656,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81771723)

摘　　要：目的探讨结直肠癌中靶向调控特定富含AT碱基DNA序列结合蛋白(SATB)2有关的miR-31和miR-182表达水平,并分析其与SATB2的关系。方法根据候选miRNAs的预测结果,构建SATB2基因3′UTR突变性和野生型的荧光素酶载体,与miRNA抑制物转染、报告基因检测和miRNA模拟物等技术手段相结合,检验候选miRNA是否对SATB2基因的表达具有靶向调控作用,确定miRNA对调控结直肠癌转移相关基因SATB2的靶效性。结果 miR-31和miR-182是靶向调控SATB2的候选基因。结直肠癌组织中miR-31和miR-182表达明显增高(P<0.05),与未转移比较,结直肠癌转移的miR-31和miR-182表达明显增高(P<0.05)。慢病毒感染结直肠癌细胞SW480得到稳定过表达。miR-31和miR-182的结直肠癌细胞株SW480/miR-31和SW480/miR-182的增殖能力明显高于对照SW480/Con(P<0.05)。SW480/miR-31和SW480/miR-182细胞形成的克隆较对照SW480/Con细胞明显多(P<0.05),SW480/miR-31,停留在G0/G1期的细胞减少,S期的细胞增多,G2/M期细胞增多,SW480/miR-182,G0/G1期的细胞减少,S期细胞增多,G2/M期细胞增多。与对照SW480/Con细胞比较,SW480/miR-31和SW480/miR-182的细胞内荧光素酶的活性明显降低(P<0.05)。SATB2基因转染后,miR-31和miR-182细胞体外增殖能力均明显降低(P<0.05)。SATB2基因转染后,SW480/miR-31和SW480/miR-182细胞进入G0/G1期,S期细胞减少,G2/M期细胞减少,SATB2基因转染后,miR-31和miR-182细胞运动能力明显降低(P<0.05)。与对照组比较,实验组裸鼠的瘤体积和瘤重量均明显增高(P<0.05)。与对照组比较,实验组裸鼠瘤的miR-31和miR-182表达均明显增高(P<0.05)。结论结直肠癌中靶向调控SATB2细胞的miR-31和miR-182在结直肠癌中的表达水平高低与癌细胞转移、克隆及预后相关,miR-31和miR-182能反向调控SATB2的表达。Objective To investigate the expressions of miR-31 and miR-182 in specific colorectal cancer targeting specific AT-rich sequence-binding protein 2(SATB2). Methods Based on the predicted results of candidate miRNAs, the 3′UTR mutation and wild-type luciferase vector of SATB2 gene were constructed and combined with miRNA inhibitor transfection, reporter gene detection and miRNA mimics to test whether candidate miRNAs were tested to determine the target effect of miRNAs on the regulation of colorectal cancer metastasis-related gene SATB2.Results miR-31 and miR-182 were candidate genes for targeted regulation of SATB2. The expressions of miR-31 and miR-182 in colorectal cancer tissues had significantly increased(P<0.05), and the expressions of miR-31 and miR-182 in colorectal cancer metastasis had significantly increased than those in non-metastasis(P<0.05). The proliferation of colorectal cancer cell lines SW480/miR-31 and W480/miR-182 stably overexpressing miR-31 and miR-182 by lentivirus-infected colorectal cancer cell SW480 had significantly increased than those of control SW480/Con(P<0.05). SW480/miR-31 and SW480/miR-182 cells formed more clones than control SW480/Con cells(P<0.05), SW480/miR-31 cells that stayed in G0 G1 phase had decreased, cells in S phase had increased, cells in G2 M phase had increased, SW480/miR-182, CRC cells in G0 G1 phase had decreased, cells in S phase had increased, and cells in G2 M phase had increased. Compared with SW480/Con cells, the activities of intracellular luciferase of SW480/miR-31 and SW480/miR-182 had significantly decreased(P<0.05). After transfection of SATB2 gene, the proliferation ability of miR-31 and miR-182 cells had significantly decreased(P<0.05). After SATB2 gene transfection, SW480/miR-31 and SW480/miR-182 cells entering G0 G1 phase, S phase cells had decreased, G2 M phase cells had decreased. After SAR2 gene transfection, miR-31 and miR-182 cells had decreased significantly(P<0.05). Compared with those of control group, the tumor volume and tumor weight of t

关 键 词：结直肠癌 特定富含AT碱基DNA序列结合蛋白2 miR-31 miR-182 

分 类 号：R34[医药卫生—基础医学]