LncRNA SNHG20通过miR-520f-3p调控胆管癌细胞增殖和侵袭

LncRNA SNHG20 regulates proliferation and invasion of cholangiocarcinoma through miR-520f-3p

目的探讨lncRNA SNHG20和miR-520f-3p对胆管癌细胞增殖和侵袭的影响及其潜在的作用机制。方法收集2012年3月至2014年3月在哈尔滨医科大学附属第二医院手术切除的20例胆管癌患者肿瘤组织及相应癌旁组织,利用脂质体转染技术分别将si-SNHG20、miR-520f-3p mimics和miR-520f-3p inhibitor及其对照质粒转染至胆管癌CCLP-1细胞,构建过表达和沉默细胞模型。利用qRT-PCR分别检测胆管癌组织和相应癌旁正常组织,以及胆管癌细胞系CCLP-1、QBC939、RBE、TFK-1和正常胆管上皮细胞系HIBEC中SNHG20与miR-520f-3p的表达水平。采用CCK-8法检测细胞增殖能力,划痕和Transwell实验分别检测细胞迁移和侵袭能力。双荧光素酶报告基因实验验证SNHG20与miR-520f-3p的靶向关系。结果分别与癌旁组织和正常胆管上皮细胞系HIBEC比较,SNHG20在胆管癌组织和胆管癌细胞系CCLP-1、QBC939、RBE、TFK-1中的表达升高(P<0.05),而miR-520f-3p表达降低(P<0.05)。与si-NC组比较,敲低SNHG20后CCLP-1细胞的增殖、迁移和侵袭能力降低(P<0.05)。SNHG20可以靶向作用并调控miR-520f-3p的表达,敲低miR-520f-3p能逆转下调SNHG20对CCLP-1细胞增殖和侵袭的抑制作用(t=10.533,P=0.002;t=8.683,P=0.037)。结论LncRNA SNHG20能靶向作用于miR-520f-3p,进而调控胆管癌细胞增殖和侵袭。

ObjectiveTo investigate the effects of lncRNA SNHG20 and miR-520 f-3 p on the proliferation,migration and invasion of cholangiocarcinoma cells and the potential mechanisms.MethodsThe tumor tissues and corresponding adjacent tissues of 20 patients with cholangiocarcinoma who were surgically resected in the 2 nd Affiliated Hospital of Harbin Medical University from March 2012 to March 2014 were collected.The si-SNHG20,miR-520 f-3 p mimics and miR-520 f-3 p inhibitor and their corresponding control plasmids were transfected into cholangiocarcinoma cells CCLP-1 by liposome transfection technology,respectively,to construct overexpression and silence cell models.The qRT-PCR was used to detect the expression levels of SNHG20 and miR-520 f-3 p in cholangiocarcinoma tissues and corresponding normal tissues as well as cholangiocarcinoma cells CCLP-1,QBC939,RBE,TFK-1 and normal bile duct epithelial cells HIBEC.The cell proliferation ability was detected by CCK-8 assay.The cell migration and invasion ability were detected by wound healing and Transwell assay,respectively.The targeting relationship between SNHG20 and miR-520 f-3 p was verified by the dual luciferase reporter gene experiment.ResultsCompared with the adjacent tissues and normal bile duct epithelial cells HIBEC,the expressions of SNHG20 in CCLP-1,QBC939,RBE,TFK-1 of cholangiocarcinoma cells were significantly higher(P<0.05),while the expressions of miR-520 f-3 p were decreased(P<0.05).Compared with si-NC group,the proliferation,migration and invasion of CCLP-1 cells were decreased after SNHG20 knockdown(P<0.05).SNHG20 could target and regulate the expression of miR-520 f-3 p,and the knockdown of miR-520 f-3 p could reverse the inhibitory effect of down-regulated SNHG20 on the proliferation and invasion of CCLP-1 cells(t=10.533,P=0.002;t=8.683,P=0.037).ConclusionsSNHG20 can target miR-520 f-3 p to regulate the proliferation and invasion of cholangiocarcinoma cells.