Cheng Hailing, Wang Ning, Tian Jun, Li Yanyun, Ren Lu, Shi Zhenyu
Department of Obstetrics and Gynecology, Huaihe Hospital of Henan University, Kaifeng, Henan, People's Republic of China.
Henan Medical School, Henan University, Kaifeng, Henan, People's Republic of China.
Cancer Manag Res. 2020 Apr 23;12:2753-2765. doi: 10.2147/CMAR.S241372. eCollection 2020.
Circular RNAs (circRNAs) are significant molecular targets in various types of human cancers. The functional mechanism of circRNA_0025033 (circ_0025033) in ovarian cancer (OC) was discussed in the current report.
The quantitative real-time polymerase chain reaction (qRT-PCR) was used for determining the circ_0025033 and microRNA-330-5p (miR-330-5p) levels. Cell Counting Kit-8 (CCK-8) and transwell assays were separately exploited to analyze cell viability and migration/invasion. Cell apoptosis was assessed using flow cytometry. The protein levels of epithelial-mesenchymal transition (EMT)-related makers and kallikrein-related peptidase 4 (KLK4) were measured by Western blotting. The target combination was confirmed by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) and RNA pull-down assays. And the effect of circ_0025033 on OC in vivo was explored via xenograft tumor assay.
Circ_0025033 was overexpressed in OC tissues and cells. Circ_0025033 knockdown inhibited OC cell viability, migration, invasion and EMT while expedited apoptosis. MiR-330-5p was a target of circ_0025033 and circ_0025033 regulated OC cellular behaviors by sequestering miR-330-5p. Moreover, miR-330-5p targeted KLK4 and circ_0025033 affected the KLK4 expression by sponging miR-330-5p. And miR-330-5p functioned as a tumor inhibitor in OC via targeting KLK4. In vivo, circ_0025033 promoted OC growth by the miR-330-5p/KLK4 axis.
This study demonstrated that circ_0025033 contributed to the progression of OC via the miR-330-5p/KLK4 axis and might be a candidate target in the identification and treatment of OC.
环状RNA(circRNAs)是各类人类癌症中的重要分子靶点。本报告探讨了circRNA_0025033(circ_0025033)在卵巢癌(OC)中的功能机制。
采用定量实时聚合酶链反应(qRT-PCR)测定circ_0025033和微小RNA-330-5p(miR-330-5p)水平。分别利用细胞计数试剂盒-8(CCK-8)和Transwell实验分析细胞活力及迁移/侵袭能力。通过流式细胞术评估细胞凋亡情况。采用蛋白质免疫印迹法检测上皮-间质转化(EMT)相关标志物和激肽释放酶相关肽酶4(KLK4)的蛋白水平。通过双荧光素酶报告基因实验、RNA免疫沉淀(RIP)实验和RNA下拉实验确认靶标结合情况。并通过异种移植瘤实验探究circ_0025033对体内OC的影响。
circ_0025033在OC组织和细胞中过表达。敲低circ_0025033可抑制OC细胞活力、迁移、侵袭及EMT,同时促进细胞凋亡。miR-330-5p是circ_0025033的靶标,circ_0025033通过结合miR-330-5p调控OC细胞行为。此外,miR-330-5p靶向KLK4,circ_0025033通过海绵吸附miR-330-5p影响KLK4表达。且miR-330-5p通过靶向KLK4在OC中发挥肿瘤抑制作用。在体内,circ_0025033通过miR-330-5p/KLK4轴促进OC生长。
本研究表明,circ_0025033通过miR-330-5p/KLK4轴促进OC进展,可能是OC诊断和治疗的候选靶点。