微小RNA-122靶向锌指和BTB结构域蛋白20调控Wnt通路影响胃癌的侵袭和迁移  

MicroRNA - 122 targets zinc finger and BTB domain containing 20 regulating the invasion and mi- gration of gastric cancer by Wnt signaling pathway

作　　者：袁远[1] 宋玉成[1] 张广坛[1] 张学东[1] 薛焕洲[1] 

机构地区：[1]郑州大学人民医院(河南省人民医院)胃肠外科,450003

出　　处：《中华实验外科杂志》2017年第4期608-611,共4页Chinese Journal of Experimental Surgery

摘　　要：目的 探讨微小RNA（miRNA，miR）-122靶向锌指和BTB结构域蛋白20（ZBTB20）调控Wnt信号通路影响胃癌的侵袭和迁移的作用机制。方法 应用实时荧光定量聚合酶链反应（FQ-PCR）检测miR-122在胃癌组织中表达；运用Western blot在胃癌细胞株中挑出高表达ZBTB20的细胞株；双荧光素酶报告基因系统检测miR-122对ZBTB20转录活性的影响；Transwell侵袭实验检测miR-122的表达对胃癌细胞SGC-7901的侵袭能力的影响；划痕实验检测miR-122的表达对胃癌细胞SGC-7901的迁移能力的影响；Western blot检测沉默ZBTB20后Wnt信号通路的蛋白表达水平。裸鼠体外成瘤实验检测过表达miR-122后SGC-7901胃癌细胞体外成瘤能力变化。 结果 miR-122在胃癌组织中低表达，ZBTB20在SGC-7901胃癌细胞株中表达水平较高（2.51±0.31比0.58±0.17, P＝0.009）；过表达miR-122后，胃癌细胞株SGC-7901的侵袭和迁移能力明显降低[（18.62±2.32）%比（35.92±2.48）%，P＝0.029；（34.25±5.75）%比（62.19±7.23）%，P＝0.032；（49.73±3.91）%比（85.24±6.29）%，P＝0.026]；双荧光素酶报告基因系统检测结果显示，miR-122可以直接调控ZBTB20的转录活性；沉默ZBTB20后，Wnt、c-Myc和细胞周期素D1（Cyclin D1）蛋白表达下调[（12.32±0.81）%比（81.45±3.69）%; （15.64±1.11）%比（88.45±8.34）%, （13.52±0.71）%比（79.45±3.76）%，P＝0.043]；过表达miR-122后，胃癌SGC-7901细胞株体外成瘤能力下降[体积（3.09±0.23） cm3比（0.45±0.12） cm3，P＝0.029；重量（2.85±0.35） g比（0.48±0.14） g，P＝0.018]。 结论miR-122靶向ZBTB20的表达，从而调控胃癌细胞的侵袭和迁移能力。Objective To investigate the effect of microRNA （miRNA, miR）-122 targeting zinc finger and BTB domain containing 20 （ZBTB20） on Wnt signaling pathway on invasion and metastasis of gastric cancer.Methods The expression of miR-122 in gastric carcinoma was detected by real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）. The expression of ZBTB20 in gastric cancer cell lines was detected by western blotting. The effect of miR-122 on the transcriptional activity of ZBTB20 was examined by the dual luciferase reporter gene system. Transwell invasion assay was used to detect the expression of miR-122 in the gastric cancer cell line SGC-7901. To detect the expression of ZBTB20 after slience miR-122 by Western blotting. Western blotting was used to detect the protein expression of Wnt signal pathway after silencing ZBTB20. The protein expression of ZBTB20 was detected by Western blotting. The ablity of vitro tumorigenesis of SGC-7901 gastric carcinoma cells after overexpressing miR-122 in nude mice was detected by tumorigenesis experiment.Results The expression of miR-122 and ZBTB20 was low in gastric cancer tissues （2.51±0.31 vs. 0.58±0.17, P=0.009）. The expression of miR-122 was significantly lower in SGC-7901 than that other cell lines. After overexpression of miR-122, the invasion and migration ability of gastric cancer cell line SGC-7901 was significantly decreased [（18.62±2.32）% vs. （35.92±2.48）%, P=0.029; （34.25±5.75）% vs. （62.19±7.23）%, P=0.032; （49.73±3.91）% vs. （85.24±6.29）%, P=0.026]. The dual luciferase reporter gene system showed that miR-122 can directly regulate the transcriptional activity of ZBTB20. After overexpression of miR-122 in gastric cancer cells. The expression level of ZBTB20 was down-regulated after silencing ZBTB20, and the expression of Wnt, c-Myc and Cyclin D1 protein were down-regulated after miR-122 overexpression [（12.32±0.81）% vs. （81.45±3.69）%; （15.64±1.11）% vs. （88.45±8.34）%, （13.52±0.71）% vs. （7

关 键 词：微小RNA-122 锌指和BTB结构域蛋白20 WNT 胃癌 

分 类 号：R735.2[医药卫生—肿瘤]