Exosome-mediated circ_0000218 targets miR-144 and regulates proliferation, migration and invasion of cholangiocarcinoma cells
渠海 1孙衍 2赵立军 1张宁 1李志强 1张翔
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作者信息
1. 1长治医学院附属和济医院普外肝胆科，长治 046011
2. 2长治医学院附属和济医院内分泌科，长治 046011
折叠
摘要
目的 探讨外泌体介导的circ_0000218对胆管癌细胞增殖、迁移和侵袭的影响及其分子机制。 方法 分离提取胆管癌细胞外泌体，qRT-PCR检测外泌体中circ_0000218表达水平，将si-circ_0000218转染至胆管癌细胞，提取外泌体分别与RBE和HuH-28细胞共培养；CCK8检测细胞活力；Transwell检测细胞迁移和侵袭；免疫印迹检测相关蛋白的表达；双荧光素酶报告基因实验检测circ_0000218和miR-144的靶向关系；功能回复实验观察circ_0000218/miR-144轴对CCA细胞增殖、迁移和侵袭的影响。 结果 circ_0000218在胆管癌组织和细胞中高表达（P<0.05）；外泌体介导的敲低circ_0000218可降低RBE和HuH-28细胞活力、迁移和侵袭数目以及Ki67、MMP-2、MMP-9表达水平（P<0.05）；circ_0000218靶向调控miR-144表达（P<0.05），且下调miR-144逆转了敲低circ_0000218表达对RBE、HuH-28细胞的增殖、迁移和侵袭的影响（P<0.05）。 结论 外泌体介导的circ_0000218可通过靶向调控miR-144表达影响胆管癌细胞的增殖、迁移和侵袭。 Objective To investigate the effect of exosome-mediated circ_0000218 on the proliferation, migration and invasion of cholangiocarcinoma cells and its molecular mechanism. Methods The exosomes were isolated and extracted from cholangiocarcinoma cells, and the expression level of circ_0000218 in the exosomes was determined by qRT-PCR. si-circ_0000218 was transfected into cholangiocarcinoma cells, and extracted exosomes were co-cultured with RBE and Huh-28 cells, respectively. Cell counting kit-8 (CCK8) assay was used to examine cell viability. Transwell assay was used to detect cell migration and invasion. Western blotting was used to measure the expression levels of related proteins. Dual luciferase reporter assay was used to identify the targeting relationship between circ_0000218 and miR-144. Functional recovery assay was used to detect the effects of circ_0000218/miR-144 axis on the proliferation, migration and invasion of CCA cells. Results circ_0000218 was highly expressed in cholangiocarcinoma tissues and cells (P<0.05). Exosome-mediated knockdown of circ_0000218 reduced the RBE and Huh-28 cell viability, migration and invasion numbers, and decreased the expression levels of Ki67, MMP-2 and MMP-9 (allP<0.05). circ_0000218 targeted and regulated the expression of miR-144 (P<0.05), and the down-regulation of miR-144 reversed the effects of knockdown of circ_0000218 expression on the proliferation, migration and invasion of RBE and Huh-28 cells (allP<0.05) . Conclusion Exosome-mediated circ_0000218 may affect proliferation, migration and invasion of cholangiocarcinoma cells by targeting and regulating the expression of miR-144.
Abstract
Objective To investigate the effect of exosome-mediated circ_0000218 on the proliferation, migration and invasion of cholangiocarcinoma cells and its molecular mechanism. Methods The exosomes were isolated and extracted from cholangiocarcinoma cells, and the expression level of circ_0000218 in the exosomes was determined by qRT-PCR. si-circ_0000218 was transfected into cholangiocarcinoma cells, and extracted exosomes were co-cultured with RBE and Huh-28 cells, respectively. Cell counting kit-8 (CCK8) assay was used to examine cell viability. Transwell assay was used to detect cell migration and invasion. Western blotting was used to measure the expression levels of related proteins. Dual luciferase reporter assay was used to identify the targeting relationship between circ_0000218 and miR-144. Functional recovery assay was used to detect the effects of circ_0000218/miR-144 axis on the proliferation, migration and invasion of CCA cells. Results circ_0000218 was highly expressed in cholangiocarcinoma tissues and cells (P<0.05). Exosome-mediated knockdown of circ_0000218 reduced the RBE and Huh-28 cell viability, migration and invasion numbers, and decreased the expression levels of Ki67, MMP-2 and MMP-9 (allP<0.05). circ_0000218 targeted and regulated the expression of miR-144 (P<0.05), and the down-regulation of miR-144 reversed the effects of knockdown of circ_0000218 expression on the proliferation, migration and invasion of RBE and Huh-28 cells (allP<0.05) . Conclusion Exosome-mediated circ_0000218 may affect proliferation, migration and invasion of cholangiocarcinoma cells by targeting and regulating the expression of miR-144.
关键词
胆管癌/外泌体/环状RNA/增殖/迁移/侵袭
Key words
Exosomes/Rabies virus/Glycoprotein/Magnetic iron oxide nanoparticles/Magnetic resonance imaging/Drug targeting/Molecular probe
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