Wang Wenshuang, Xu Anli, Zhao Manyin, Sun Jianan, Gao Lingyun
Department of Gynecology and Obstetrics, Yantai Yuhuangding Hospital, Qingdao University, Yantai, 264000, Shandong, China.
Department of Ultrasound, Yantai Yuhuangding Hospital, Qingdao University, NO.20, East Yuhuangding Road, Yantai, 264000, Shandong, China.
Biotechnol Lett. 2021 Mar;43(3):745-755. doi: 10.1007/s10529-020-03059-w. Epub 2021 Jan 1.
There is increasing evidence that circular RNA (circRNA) disorders have an impact on the progression of various malignancies. The expression characteristics, function and underlying mechanism of circ_0001247 in cervical cancer (CC) have not been confirmed.
GSE147483 datasets of circRNAs expression in CC cell line and normal cervical cell line were retrieved from GEO database, and the circRNA with significant difference was selected; circ_0001247, miR-1270, and Zinc finger E-box binding homeobox 2 (ZEB2) expressions in CC tissues and cell lines were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) assay; cell counting kit-8 (CCK-8) assay and BrdU assay were applied to monitor the proliferative ability of CC cells; Transwell assay was conducted to examine the migration and invasion of CC cells, and flow cytometry was used to evaluate the apoptosis; Western blot assay was adopted to detect ZEB2 protein expressions; dual-luciferase report gene assay was used to verify the targeting relationship between circ_0001247 and miR-1270, and miR-1270 and the 3'UTR of ZEB2.
Analysis of GSE147483 suggested that circ_0001247 could probably be an oncogenic circRNA in CC. Compared with that in adjacent tissues and normal cervical epithelial cells, circ_0001247 expression in CC tissues and cell lines was significantly increased; knocking down circ_0001247 expression could inhibit the proliferation and metastasis of CC cells, and promote apoptosis, while circ_0001247 overexpression worked oppositely; circ_0001247 sponged miR-1270 in CC cells; miR-1270 diminished the promoting effect of circ_0001247 by inactivating the ZEB2.
Circ_0001247 promotes progression of CC by sponging miR-1270 to upregulate ZEB2 expression level.
越来越多的证据表明,环状RNA(circRNA)紊乱对各种恶性肿瘤的进展有影响。circ_0001247在宫颈癌(CC)中的表达特征、功能及潜在机制尚未得到证实。
从基因表达综合数据库(GEO数据库)中检索CC细胞系和正常宫颈细胞系中circRNA表达的GSE147483数据集,并选择差异显著的circRNA;采用定量实时聚合酶链反应(qRT-PCR)检测CC组织和细胞系中circ_0001247、微小RNA-1270(miR-1270)和锌指E盒结合同源框2(ZEB2)的表达;应用细胞计数试剂盒-8(CCK-8)检测和5-溴脱氧尿嘧啶核苷(BrdU)检测监测CC细胞的增殖能力;采用Transwell检测分析CC细胞的迁移和侵袭能力,并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(Western blot)检测ZEB2蛋白表达;采用双荧光素酶报告基因检测验证circ_0001247与miR-1270以及miR-1270与ZEB2的3'非翻译区(3'UTR)之间的靶向关系。
对GSE147483的分析表明,circ_0001247可能是CC中的一种致癌circRNA。与相邻组织和正常宫颈上皮细胞相比,circ_0001247在CC组织和细胞系中的表达显著增加;敲低circ_0001247表达可抑制CC细胞的增殖和转移,并促进细胞凋亡,而circ_0001247过表达则产生相反的作用;circ_0001247在CC细胞中可吸附miR-1270;miR-1270通过使ZEB2失活减弱circ_0001247的促进作用。
Circ_0001247通过吸附miR-1270上调ZEB2表达水平促进CC进展。