Hsa_circ_001680 affects the proliferation and migration of CRC and mediates its chemoresistance by regulating BMI1 through miR-340
Hsa_circ_001680 affects the proliferation and migration of CRC and mediates its chemoresistance by regulating BMI1 through miR-340
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Hsa_circ_001680通过miR-340调节BMI1影响CRC的增殖和迁移并介导其化疗耐药
DOI：
10.1186/s12943-020-1134-8
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发表时间：
2020-01-31
期刊：
影响因子：
37.3
通讯作者：
Li, Tingting
中科院分区：
文献类型：
--
作者：
Jian, Xiangyu;He, Han;Li, Tingting
背景越来越多的证据表明，环状RNA（circRNA）作为微小RNA（miRNA）海绵，直接抑制特定的miRNA并改变其在转录后水平调节基因表达的能力；据信这种机制发生在各种癌症中。然而，circ_001680在结直肠癌（CRC）中的表达水平、确切功能和机制尚不清楚。方法采用qRT-PCR检测人CRC组织及其匹配的正常组织中circ_001680和miR-340的表达。使用生物信息学分析和双荧光报告基因测定来评估circ_001680是否可以与miR-340结合。构建了 Circ_001680 过表达和敲低细胞系，通过基于功能的实验（包括 CCK8、板克隆形成、Transwell 和伤口愈合测定）研究体内和体外的增殖和迁移能力。通过生物信息学分析、双荧光报告系统、FISH、RIP 和 RNA pull down 分析研究了 circ_001680、miR-340 和 BMI1 之间的关系。采用球形成实验和流式细胞术分析评估circ_001680对CRC细胞干性特征的影响。结果Circ_001680在CRC组织中的表达高于来自同一患者的匹配的邻近正常组织。观察到 Circ_001680 可以增强 CRC 细胞的增殖和迁移能力。此外，双荧光报告基因检测证实，circ_001680 通过靶向 miR-340 影响 BMI1 的表达。更重要的是，我们还发现circ_001680可以促进CRC中的癌症干细胞（CSC）群体，并通过调节miR-340靶基因BMI1诱导伊立替康治疗耐药。结论我们的结果表明，circ_001680是通过BMI1上调诱导CRC化疗耐药的新策略的一部分。此外，circ_001680可能是一种有前途的诊断和预后标志物，用于确定基于伊立替康的化疗是否成功。
BackgroundAccumulating evidence indicates that circular RNAs (circRNAs) act as microRNA (miRNA) sponges to directly inhibit specific miRNAs and alter their ability to regulate gene expression at the post-transcriptional level; this mechanism is believed to occur in various cancers. However, the expression level, precise function and mechanism of circ_001680 in colorectal carcinoma (CRC) are largely unknown.MethodsqRT-PCR was used to detect the expression of circ_001680 and miR-340 in human CRC tissues and their matched normal tissues. Bioinformatics analyses and dual-fluorescence reporter assays were used to evaluate whether circ_001680 could bind to miR-340. Circ_001680 overexpression and knockdown cell lines were constructed to investigate the proliferation and migration abilities in vivo and in vitro through function-based experiments, including CCK8, plate clone formation, transwell, and wounding healing assays. The relationships among circ_001680, miR-340 and BMI1 were investigated by bioinformatics analyses, dual-fluorescence reporter system, FISH, RIP and RNA pull down assays. Sphere forming assays and flow cytometry analyses were used to assess the effect of circ_001680 on the stemness characteristics of CRC cells.ResultsCirc_001680 was more highly expressed in of CRC tissue than in matched adjacent normal tissues from the same patients. Circ_001680 was observed to enhance the proliferation and migration capacity of CRC cells. Furthermore, dual-fluorescence reporter assays confirmed that circ_001680 affects the expression of BMI1 by targeting miR-340. More importantly, we also found that circ_001680 could promote the cancer stem cell (CSC) population in CRC and induce irinotecan therapeutic resistance by regulating the miR-340 target gene BMI1.ConclusionsOur results demonstrated that circ_001680 is a part of a novel strategy to induce chemotherapy resistance in CRC through BMI1 upregulation. Moreover, circ_001680 may be a promising diagnostic and prognostic marker to determine the success of irinotecan-based chemotherapy.