circNT5E通过调控miR-506-3p表达对结肠癌细胞迁移、侵袭和凋亡的影响  

Effect of circNT5E on the migration,invasion and apoptosis of colon cancer cells by regulating the expression of miR-506-3p

作　　者：孟凡迪[1] 徐勤鸿[1] 杨刚华[1] 万永[1] 张雷[1] MENG Fandi;XU Qinhong;YANG Ganghua;WAN Yong;ZHANG Lei(Department of geriatric surgery,the first affiliated hospital of xi'an jiaotong university,Xi'an,Shanxi,China,710061)

机构地区：[1]西安交通大学第一附属医院老年外科,陕西西安710061

出　　处：《分子诊断与治疗杂志》2020年第6期760-764,共5页Journal of Molecular Diagnostics and Therapy

基　　金：陕西省科技攻关项目(2019SF-157、2018SF-055)。

摘　　要：目的探讨circNT5E对结肠癌细胞迁移、侵袭和凋亡的影响及分子机制。方法选取40例2017年1月至2019年5月本院结肠癌患者癌组织及其相应癌旁组织;将结肠癌细胞HCT116分为si-NC组、si-circNT5E组、pcDNA组、pcDNA-circNT5E组、miR-NC组、miR-506-3p组、si-circNT5E+antimiR-NC组、si-circNT5E+anti-miR-506-3p组。实时荧光定量PCR(RT-qPCR)检测circNT5E和miR-506-3p的表达水平;显色原位杂交(CISH)检测癌组织中circNT5E表达;Transwell检测细胞迁移和侵袭;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达;双荧光素酶报告实验验证circNT5E和miR-506-3p的靶向关系。结果与癌旁组织相比,结肠癌组织中circNT5E表达水平显著升高,miR-506-3p表达水平显著降低(P<0.05)。沉默circNT5E或过表达miR-506-3p,迁移、侵袭细胞数显著降低,细胞凋亡率显著升高,MMP-2、MMP-9、Bcl-2表达水平显著降低,Bax表达水平显著升高(P<0.05)。circNT5E靶向调控miR-506-3p,抑制miR-506-3p表达逆转了沉默circNT5E表达对结肠癌HCT116细胞迁移、侵袭和凋亡的作用。结论沉默circNT5E表达可能通过上调miR-506-3p的表达抑制结肠癌HCT116细胞迁移、侵袭,促进细胞凋亡。Objective To investigate the effect and molecular mechanism of circNT5E on colon cancer cell migration,invasion and apoptosis.Methods 40 cases of colon cancer patients tissues and their corresponding adjacent tissues from January 2017 to May 2019 from our hospital were selected.The colon cancer cells(HCT116)were divided into si-NC group,si-circNT5E group,pcDNA group,pcDNA-circNT5E group,miR-NC group,miR-506-3p group,si-circNT5E+anti-miR-NC group,si-circNT5E+anti-miR-506-3p group.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expressions of circNT5E and miR-506-3p.Color in situ hybridization(CISH)was used to detect the expression of circNT5E in cancer tissues.Transwell was used to detect cell migration and invasion Flow cytometry was used to detect apoptosis.Western blot was used to detect matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9),B-cell lymphoma/leukemia-2(Bcl-2),Bcl-2 related X protein(Bax)expression.Dual luciferase report assay was used to target the relationship between circNT5E and miR-506-3p.Results Compared with adjacent cancer tissues,the expression of circNT5E in colon cancer tissues was significantly increased,and the expression of miR-506-3p was significantly decreased(P<0.05).Silence of circNT5E or overexpression of miR-506-3p,the number of migrating and invading cells was significantly reduced,the apoptosis rate was significantly increased,the expressions of MMP-2,MMP-9 and Bcl-2 were significantly reduced,and Bax expression was significantly increased(P<0.05).circNT5E targets regulation of miR-506-3p,and inhibits the miR-506-3p expression reversed the effects of silent circNT5E expression on the migration,invasion and apoptosis of HCT116 cells.Conclusions Silencing circNT5E expression inhibits the migration and invasion of HCT116 cells and promotes apoptosis by up-regulating the expression of miR-506-3p.

关 键 词：circNT5E miR-506-3p 结肠癌 迁移 侵袭 凋亡 

分 类 号：R735.35[医药卫生—肿瘤]