Yue Meng, Liu Yanxia, Zuo Taiyang, Jiang Yakun, Pan Jianmei, Zhang Shuhong, Shen Xingjie
Department of Gastroenterology, Jinan Central Hospital Affiliated to Shandong First Medical University, No.105 Jiefang Road, Lixia District, Jinan, 250013, Shandong Province, China.
Department of Oncology, Shengli Oil Central Hospital, Dongying City, Shandong Province, China.
Dig Dis Sci. 2022 Feb;67(2):492-503. doi: 10.1007/s10620-021-06894-7. Epub 2021 Feb 25.
Circular RNAs (circRNAs) can act as promoters or inhibitors in cancer progression. Has_circ_0006948 (circ_0006948) was reported to aggravate the malignant behaviors of esophageal carcinoma (EC).
This study focused on investigating the molecular mechanism of circ_0006948 in EC progression.
The quantitative real-time polymerase chain reaction was performed to detect the expression of circ_0006948, microRNA-4262 (miR-4262) and fibronectin type III domain containing 3B (FNDC3B). Cell growth analysis was conducted by Cell Counting Kit-8 and colony formation assays. Cell migration and invasion were assessed by transwell assay. Epithelial-mesenchymal transition (EMT)-associated proteins and FNDC3B protein expression were assayed using western blot. Dual-luciferase reporter and RNA pull-down assays were performed to validate the target combination. Xenograft tumor assay was used for investigating the role of circ_0006948 in vivo.
Circ_0006948 was upregulated in EC tissues and cells. Downregulating the expression of circ_0006948 or FNDC3B repressed cell growth, migration, invasion and EMT in EC cells. Target analysis indicated that miR-4262 was a target for circ_0006948 and FNDC3B was a downstream gene for miR-4262. Moreover, circ_0006948 could affect the expression of FNDC3B via sponging miR-4262. The effects of si-circ_0006948#1 on EC cells were partly restored by miR-4262 inhibition or FNDC3B overexpression. In addition, circ_0006948 also facilitated EC tumorigenesis in vivo by targeting the miR-4262/FNDC3B axis.
Taken together, circ_0006948 functioned as an oncogenic factor in EC by the miR-4262-mediated FNDC3B expression regulation.
环状RNA(circRNA)在癌症进展中可作为促进因子或抑制因子。据报道,Has_circ_0006948(circ_0006948)可加剧食管癌(EC)的恶性行为。
本研究聚焦于探究circ_0006948在EC进展中的分子机制。
采用定量实时聚合酶链反应检测circ_0006948、微小RNA-4262(miR-4262)和含III型纤连蛋白结构域3B(FNDC3B)的表达。通过细胞计数试剂盒-8和集落形成试验进行细胞生长分析。采用Transwell试验评估细胞迁移和侵袭能力。使用蛋白质免疫印迹法检测上皮-间质转化(EMT)相关蛋白和FNDC3B蛋白表达。进行双荧光素酶报告基因和RNA下拉试验以验证靶标结合情况。采用异种移植瘤试验研究circ_0006948在体内的作用。
circ_0006948在EC组织和细胞中上调。下调circ_0006948或FNDC3B的表达可抑制EC细胞的生长、迁移、侵袭和EMT。靶标分析表明,miR-4262是circ_0006948的靶标,FNDC3B是miR-4262的下游基因。此外,circ_0006948可通过海绵吸附miR-4262影响FNDC3B的表达。抑制miR-4262或过表达FNDC3B可部分恢复si-circ_0006948#1对EC细胞的作用。此外,circ_0006948还通过靶向miR-4262/FNDC3B轴促进EC在体内的肿瘤发生。
综上所述,circ_0006948通过miR-4262介导的FNDC3B表达调控在EC中发挥致癌因子的作用。