Department of Cell-Gene Therapy Translational Medicine Research Center, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
Department of General Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
J Cell Physiol. 2021 Feb;236(2):1252-1269. doi: 10.1002/jcp.29931. Epub 2020 Jul 15.
Circular RNAs (circRNA) have been reported as regulators involved in hepatocellular carcinoma (HCC), but their mechanism of activity remains unknown. This study performed quantitative reverse-transcription polymerase chain reaction to determine if circNFATC3 was downregulated in 46 paired HCC tissues and cell lines. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, apoptotic, and transwell assay proved that circNFATC3 can inhibit hepatoma cell proliferation, apoptosis, and migration/invasion in vitro. Mouse xenograft assay demonstrated that circNFATC3 suppressed tumor size and weight and reduced lung metastasis in vivo, and vice versa. The RNA-seq results showed that NFATC3 itself was the most significantly differentially expressed gene when circNFATC3 was manipulated, and bioinformatics and luciferase reporter assays verified circNFATC3 regulated the expression of NFATC3 by interacting with the hsa-miR-548I. Additionally, it was also indicated that the level of NFATC3 was downregulated in HCC patients also and was significantly correlated with the staging and prognosis of HCC. Moreover, both circNFATC3 and NFATC3 were shown to inhibit the phosphorylation of JNK, c-Jun, AKT, and mTOR signaling pathways. Overall, the circNFATC3 can sponge miR-548I to protect NFATC3 itself, then it regulates hepatoma cell function via the JNK, c-Jun, AKT, and mTOR signaling pathways, and the circNFATC3 can be a tumor-repressor on HCC.
环状 RNA（circRNA）已被报道为参与肝细胞癌（HCC）的调节因子，但它们的作用机制尚不清楚。本研究通过定量逆转录聚合酶链反应来确定 circNFATC3 在 46 对 HCC 组织和细胞系中是否下调。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐、凋亡和transwell 测定证明 circNFATC3 可以抑制肝癌细胞在体外的增殖、凋亡和迁移/侵袭。小鼠异种移植实验表明，circNFATC3 在体内抑制肿瘤大小和重量，并减少肺转移，反之亦然。RNA-seq 结果表明，当操纵 circNFATC3 时，NFATC3 本身是最显著差异表达的基因，生物信息学和荧光素酶报告基因实验验证了 circNFATC3 通过与 hsa-miR-548I 相互作用来调节 NFATC3 的表达。此外，还表明 NFATC3 在 HCC 患者中的水平也下调，并且与 HCC 的分期和预后显著相关。此外，circNFATC3 和 NFATC3 都被证明可以抑制 JNK、c-Jun、AKT 和 mTOR 信号通路的磷酸化。总的来说，circNFATC3 可以通过海绵吸附 miR-548I 来保护 NFATC3 本身，然后通过 JNK、c-Jun、AKT 和 mTOR 信号通路调节肝癌细胞的功能，并且 circNFATC3 可以作为 HCC 的肿瘤抑制因子。
