Ni Chuangye, Yang Shikun, Ji Yang, Duan Yunfei, Yang Wenjie, Yang Xinchen, Li Min, Xie Jun, Zhang Chuanyong, Lu Yunjie, Lu Hao
Hepatobiliary/Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University; Key Laboratory of Liver Transplantation, Chinese Academy of Medical Sciences, Nanjing, 210029, China.
Department of Hepatobiliary Surgery, The First People's Hospital of Changzhou, The Third Hospital Affiliated to Soochow University, Changzhou, 213000, China.
Cell Death Discov. 2021 Oct 1;7(1):270. doi: 10.1038/s41420-021-00664-0.
Circular RNAs (circRNAs), continuous loops of single-stranded RNA, regulate gene expression during the development of various cancers. However, the function of circRNAs in hepatocellular carcinoma (HCC) is rarely discussed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the mRNA levels of circ_0011385, miR-361-3p, and STC2 in 96 pairs of HCC tissues (tumor tissues and adjacent normal tissues), HCC cell lines, and L02 (human normal liver cell line) cells. The relationships between circ_0011385 expression and clinical features of HCC were evaluated. Functional experiments in vitro or in vivo were used to evaluate the biological function of circ_0011385. Bioinformatics analysis was performed to predict miRNAs and mRNAs sponged by circ_0011385. RNA immunoprecipitation (RIP) and dual-luciferase reporter gene assays were used to elucidate the interactions among circ_0011385, miR-361-3p, and STC2 (stanniocalcin 2). ChIP and dual-luciferase reporter gene assays were used to identify the upstream regulator of circ_0011385. High expression of circ_0011385 was observed in HCC tissues and cell lines and was significantly associated with tumor size, TNM stage, and prognosis. In addition, inhibition of circ_0011385 expression prevented the proliferation of HCC cells in vitro and in vivo. Circ_0011385 sponged miR-361-3p, thereby regulating the mRNA expression of STC2. In addition, the transcription of circ_0011385 was regulated by SP3. Circ_0011385 knockdown suppressed cell proliferation and tumor activity in HCC. Circ_0011385 may therefore serve as a new biomarker in the diagnosis and treatment of HCC.
环状RNA(circRNAs)是单链RNA的连续环,在各种癌症的发展过程中调节基因表达。然而,circRNAs在肝细胞癌(HCC)中的功能鲜有讨论。采用定量实时聚合酶链反应(qRT-PCR)检测96对HCC组织(肿瘤组织和癌旁正常组织)、HCC细胞系及L02(人正常肝细胞系)细胞中circ_0011385、miR-361-3p和STC2(甲状旁腺激素相关蛋白)的mRNA水平。评估circ_0011385表达与HCC临床特征之间的关系。采用体外或体内功能实验评估circ_0011385的生物学功能。进行生物信息学分析以预测被circ_0011385海绵化的miRNA和mRNA。采用RNA免疫沉淀(RIP)和双荧光素酶报告基因检测来阐明circ_0011385、miR-361-3p和STC2之间的相互作用。采用染色质免疫沉淀(ChIP)和双荧光素酶报告基因检测来鉴定circ_0011385的上游调节因子。在HCC组织和细胞系中观察到circ_0011385高表达,且与肿瘤大小、TNM分期及预后显著相关。此外,抑制circ_0011385表达可阻止HCC细胞在体外和体内的增殖。Circ_0011385海绵化miR-361-3p,从而调节STC2的mRNA表达。此外,circ_0011385的转录受SP3调控。Circ_0011385敲低可抑制HCC细胞增殖和肿瘤活性。因此,Circ_0011385可能成为HCC诊断和治疗的新生物标志物。