Ren Ruizhen, Du Yuanna, Niu Xing, Zang Rukun
Department of Endocrinology, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai 264000, Shandong, China.
Department of Radiotherapy, Yantai Yuhuangding Hospital Affiliated to Qingdao University, Yantai 264000, Shandong, China.
J Cancer. 2021 Apr 17;12(11):3393-3406. doi: 10.7150/jca.51437. eCollection 2021.
Our purpose was to study the roles and molecular mechanisms of long non-coding RNA (lncRNA) ZFPM2 Antisense RNA 1 (ZFPM2-AS1) in thyroid cancer. Firstly, the expression of ZFPM2-AS1, miR-515-5p and TUSC3 was detected in thyroid cancer tissues and cells. Secondary, their biological functions (proliferation, apoptosis, migration and invasion) were analyzed by a serious of functional experiments including cell counting kit-8 (CCK-8), clone formation, 5-Ethynyl-2'-deoxyuridine (EdU), enzyme-linked immunosorbent assay (ELISA), wound healing and Transwell assays. Thirdly, the mechanisms of STAT1/ZFPM2-AS1 and ZFPM2-AS1/miR-515-5p/TUSC were validated using chromatin immunoprecipitation (CHIP), pull-down and luciferase reporter assays. ZFPM2-AS1 and TUSC were both highly expressed and miR-515-5p was down-regulated in thyroid cancer tissues as well as cells. Their knockdown weakened thyroid cancer cell growth, migration, and invasion. ZFPM2-AS1 was mainly distributed in the nucleus and cytoplasm of thyroid cancer cells. Mechanistically, up-regulation of ZFPM2-AS1 was induced by transcription factor STAT1 in line with CHIP and luciferase reporter assays. Furthermore, as a sponge of miR-515-5p, ZFPM2-AS1 decreased the ability of miR-515-5p to inhibit TUSC3 expression by pull-down, luciferase reporter and gain-and-loss assays, thereby promoting malignant progression of thyroid cancer. ZFPM2-AS1 acted as an oncogene in thyroid cancer, which was transcriptionally mediated by STAT1. Furthermore, ZFPM2-AS1 weakened the inhibitory effect of miR-515-5p on TUSC3. Thus, ZFPM2-AS1 could be an underlying biomarker for thyroid cancer.
我们的目的是研究长链非编码RNA（lncRNA）锌指蛋白2反义RNA 1（ZFPM2-AS1）在甲状腺癌中的作用及分子机制。首先，检测甲状腺癌组织和细胞中ZFPM2-AS1、miR-515-5p和TUSC3的表达。其次，通过一系列功能实验，包括细胞计数试剂盒-8（CCK-8）、克隆形成、5-乙炔基-2'-脱氧尿苷（EdU）、酶联免疫吸附测定（ELISA）、伤口愈合和Transwell实验，分析它们的生物学功能（增殖、凋亡、迁移和侵袭）。第三，使用染色质免疫沉淀（CHIP）、下拉和荧光素酶报告基因实验验证STAT1/ZFPM2-AS1和ZFPM2-AS1/miR-515-5p/TUSC的机制。ZFPM2-AS1和TUSC在甲状腺癌组织和细胞中均高表达，而miR-515-5p下调。它们的敲低减弱了甲状腺癌细胞的生长、迁移和侵袭。ZFPM2-AS1主要分布在甲状腺癌细胞的细胞核和细胞质中。机制上，根据CHIP和荧光素酶报告基因实验，转录因子STAT1诱导ZFPM2-AS1上调。此外，作为miR-515-5p的海绵，ZFPM2-AS1通过下拉、荧光素酶报告基因和得失实验降低了miR-515-5p抑制TUSC3表达的能力，从而促进甲状腺癌的恶性进展。ZFPM2-AS1在甲状腺癌中作为癌基因发挥作用，由STAT1转录介导。此外，ZFPM2-AS1减弱了miR-515-5p对TUSC3的抑制作用。因此，ZFPM2-AS1可能是甲状腺癌的潜在生物标志物。
