circ_0000615通过靶向miR-432-5p调控胆管癌细胞的增殖、迁移和侵袭  

circ_0000615 regulates proliferation, migration and invasion of cholangiocarcinoma cells by targeting miR-432-5p

作　　者：关沧海 王海存 于少博 高欣 姜兴明[1] 孙东升[1] GUAN Canghai;WANG Haicun;YU Shaobo;GAO Xin;JIANG Xingming;SUN Dongsheng(Department of General Surgery,the Second Afiliated Hospital of Harbin Medical University,Harbin 150086,Heilongjiang,China;Key Laboratory of Myocardial Ischemia,the Second Afiliated Hospital of Harbin Medical University,Harbin 150086,Heilongjiang,China)

机构地区：[1]哈尔滨医科大学附属第二医院普通外科,黑龙江哈尔滨150086 [2]哈尔滨医科大学附属第二医院心肌缺血重点实验室,黑龙江哈尔滨150086

出　　处：《中国肿瘤生物治疗杂志》2022年第6期527-533,共7页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81602088);黑龙江省自然科学基金资助项目(No.LH2020H058);湖北陈孝平科技发展基金资助项目(No.CXPJJH12000002-2020015)。

摘　　要：目的:探讨circ_0000615在胆管癌细胞中的表达及其对胆管癌细胞增殖、迁移和侵袭能力的影响和可能的调控机制。方法:通过qPCR检测circ_0000615在正常胆管上皮HIBEC细胞和胆管癌CCLP-1、QBC939、TFK-1和RBE细胞中的表达水平。双荧光素酶报告基因实验验证circ_0000615与miR-432-5p的靶向关系。将si-NC、si-circ_0000615(si-circ-1、si-circ-2)、inhibitor-NC和inhibitor-miR-432-5p转染至CCLP-1和QBC939细胞,转染细胞分为si-NC组、si-circ-1组、si-circ-2组、si-NC+inh-NC组、si-NC+inh-miR-432-5p组和si-circ-1+inh-miR-432-5p组,通过CCK-8、EdU和Transwell实验检测各转染组胆管癌细胞的增殖、迁移和侵袭能力。结果:在胆管癌细胞中circ_0000615呈高表达而miR-432-5p呈低表达(P<0.05或P<0.01);双荧光素酶报告基因实验证实circ_0000615与miR-432-5p之间存在靶向关系(P<0.01);敲减circ_0000615可明显抑制CCLP-1、QBC939细胞的增殖、侵袭和迁移能力(P<0.05或P<0.01);下调miR-432-5p可部分逆转敲减circ_0000615对胆管癌CCLP-1和QBC939细胞增殖、迁移和侵袭的抑制作用(P<0.05或P<0.01)。结论:circ_0000615通过靶向miR-432-5p调控胆管癌细胞的增殖、侵袭和迁移。Objective: To investigate the expression of circular circ_0000615 in cholangiocarcinoma cells and its effect on the proliferation, migration and invasion ability of tumor cells as well as the possible regulatory mechanism. Methods: The expression level of circ_0000615 in cholangiocarcinoma cells(TFK-1, RBE, CCLP-1 and QBC939) and normal bile duct epithelium HIBEC cells was detected by qPCR. The targeting relationship between circ_0000615 and miR-432-5p was verified by the Dual-luciferase reporter gene assay. si-NC, si-circ_0000615(si-circ-1, si-circ-2), inhibitor-NC and inhibitor-miR-432-5p were separately or jointly transfected into CCLP-1 and QBC939 cells, namely si-NC group, si-circ-1 group, si-circ-2 group, si-NC+inh-NC group, si-NC+inh-miR-432-5p group, and si-circ-1+inh-miR-432-5p group. The proliferation, migration and invasion of cholangiocarcinoma cells in each group were detected by CCK-8, EdU and Transwell experiments. Results: circ_0000615 was highly expressed while miR-432-5p was lowly expressed in cholangiocarcinoma tumor cells(P<0.05 or P<0.01). Dual-luciferase reporter gene assay confirmed that there was a targeting relationship between circ_0000615 and miR-432-5p(P<0.01). Silencing circ_0000615 significantly inhibited the proliferation,invasion, and migration ability of CCLP-1 and QBC939 cells(P<0.05 or P<0.01). Silencing miR-432-5p partially reversed the inhibitory effects of circ_0000615 knockdown on proliferation, migration and invasion of cholangiocarcinoma CCLP-1 and QBC939 cells(P<0.05 or P<0.01). Conclusion: circ_0000615 regulates the proliferation, invasive and migration of cholangiocarcinoma by targeting miR-432-5p.

关 键 词：胆管癌 CCLP-1细胞 QBC939细胞 环状RNA_0000615 miR-432-5p 增殖 迁移 侵袭 

分 类 号：R735.8[医药卫生—肿瘤] R730.2[医药卫生—临床医学]