Department of Obstetrics and Gynecology, Taizhou Central Hospital (Taizhou University Hospital), Taizhou City, Zhejiang Province, China.
Anticancer Drugs. 2022 Jul 1;33(6):539-552. doi: 10.1097/CAD.0000000000001278. Epub 2022 Apr 5.
Circular RNAs (circRNAs) have a crucial role in the occurrence of many diseases, such as tumors. Yet the roles of circ_0009035 (circRACGAP1) in cervical cancer are not fully characterized. The expression levels of circRACGAP1, miR-1305 and cAMP-responsive element-binding protein 3 regulatory factor (CREBRF) were detected by using real-time quantitative PCR or western blot. Cell counting kit-8 assay, 5-ethynyl-2'-deoxyuridine, colony formation assay, transwell assay and tube formation assay were used to detect cell proliferation, migration and invasion and angiogenesis, respectively. Flow cytometry assay was used to analyze the cell apoptosis. Dual-luciferase reporter assay and RNA immunoprecipitation assay were performed to analyze the targeting about miR-1305 and circ_0009035 or CREBRF. Xenograft model was built to study the role of circ_0009035 in vivo. Immunohistochemistry was used to detect the expression of Ki67, epithelial cadherin and vimentin. First, we found that circ_0009035 expression was significantly upregulated in tumor cells and tissues; second, knockdown of circ_0009035 could inhibit cell proliferation, migration and invasion and promote cell apoptosis. Subsequently, circ_0009035 was found to be able to target miR-1305, and the expression of miR-1305 in tumor tissues and cells was significantly lower. MiR-1305 inhibitor could restore cell-related progression of cervical cancer inhibited by si-circ_0009035. Finally, miR-1305 could target CREBRF, and circ_0009035 could regulate CREBRF expression by targeting miR-1305, thereby affecting cervical cancer tumorigenesis. In summary, our study confirmed that circ_0009035 could influence the development of cervical cancer through the targeted regulation of miR-1305/CREBRF.
环状 RNA(circRNAs)在许多疾病的发生中起着关键作用,如肿瘤。然而,circ_0009035(circRACGAP1)在宫颈癌中的作用尚未完全阐明。采用实时定量 PCR 或 Western blot 检测 circRACGAP1、miR-1305 和环磷酸腺苷反应元件结合蛋白 3 调节因子(CREBRF)的表达水平。细胞计数试剂盒-8 检测、5-乙炔基-2'-脱氧尿苷、集落形成检测、Transwell 检测和管形成检测分别用于检测细胞增殖、迁移和侵袭以及血管生成。流式细胞术检测细胞凋亡。双荧光素酶报告基因检测和 RNA 免疫沉淀检测分析 miR-1305 和 circ_0009035 或 CREBRF 的靶向关系。建立异种移植模型研究 circ_0009035 在体内的作用。免疫组化检测 Ki67、上皮钙黏蛋白和波形蛋白的表达。首先,我们发现 circ_0009035 在肿瘤细胞和组织中的表达明显上调;其次,circ_0009035 的敲低可抑制细胞增殖、迁移和侵袭,促进细胞凋亡。随后发现 circ_0009035 能够靶向 miR-1305,肿瘤组织和细胞中 miR-1305 的表达明显降低。miR-1305 抑制剂可恢复 si-circ_0009035 抑制的宫颈癌细胞相关进展。最后,miR-1305 可以靶向 CREBRF,circ_0009035 可以通过靶向 miR-1305 调节 CREBRF 的表达,从而影响宫颈癌的发生。综上所述,我们的研究证实,circ_0009035 可以通过靶向调控 miR-1305/CREBRF 影响宫颈癌的发展。