Jia Benli, Yin Xiaoqiang, Wang Yong, Qian Jin, He Yan, Yang Chuang, Yu Gang, Guo Bing, Meng Xiangling
Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, People's Republic of China.
Department of General Surgery, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, People's Republic of China.
Onco Targets Ther. 2020 May 29;13:4893-4903. doi: 10.2147/OTT.S251300. eCollection 2020.
Mounting evidences reveal that circular RNAs (circRNAs) are critical to regulate biological behavior and process of tumor. Our objective is to explore the role of circRNA-PTN (circPTN) and explain the exact mechanism in hepatocellular carcinoma (HCC).
Real-time polymerase chain reaction assay was used to detect the level of circPTN and miR-326. The proliferation of cell was measured by CCK-8 assay and EdU assay. Western blot assay was performed to assess ErbB/PI3K expression. Luciferase and RNA pull-down assays were carried out to confirm the interaction between circPTN and miR-326.
Our results indicated that circPTN was upregulated in human hepatocellular carcinoma tumor tissues and cell lines, compared with paratumor tissues and immortalized normal liver cell line. circPTN could significantly promote HCC tumor growth according to gain-and loss-of-function assays. Additionally, we determined that circPTN acted as a sponge through interacting with miR-326. Overexpression of miR-326 could rescue the cell proliferation inhibition and ErbB/PI3K downregulation in HCC cells by circPTN. Besides, the effects of miR-326 on HCC were missing when circPTN binding sites were mutated.
Our study indicates that circPTN acts as an oncogenic factor via sponging miR-326 in HCC.
越来越多的证据表明,环状RNA(circRNA)对调节肿瘤的生物学行为和进程至关重要。我们的目的是探讨环状RNA-PTN(circPTN)的作用,并阐明其在肝细胞癌(HCC)中的具体机制。
采用实时聚合酶链反应检测circPTN和miR-326的水平。通过CCK-8法和EdU法检测细胞增殖情况。采用蛋白质免疫印迹法评估ErbB/PI3K的表达。进行荧光素酶报告基因实验和RNA下拉实验以证实circPTN与miR-326之间的相互作用。
我们的结果表明,与癌旁组织和永生化正常肝细胞系相比,circPTN在人肝细胞癌肿瘤组织和细胞系中上调。根据功能获得和功能缺失实验,circPTN可显著促进肝癌肿瘤生长。此外,我们确定circPTN通过与miR-326相互作用发挥海绵作用。miR-326的过表达可挽救circPTN对肝癌细胞增殖的抑制作用以及ErbB/PI3K的下调。此外,当circPTN结合位点发生突变时,miR-326对肝癌的作用消失。
我们的研究表明,circPTN在肝癌中通过充当miR-326的海绵发挥致癌因子的作用。