Department of Pathology, The First Affiliated Hospital of NanChang University, Nanchang, China.
Department of General Thyroid Surgery, The First Affiliated Hospital of NanChang University, Nanchang, China.
Biosci Rep. 2021 Mar 26;41(3). doi: 10.1042/BSR20201603.
Long non-coding RNAs (lncRNAs) recently have been identified as influential indicators in a variety of malignancies. The aim of the present study was to identify a functional lncRNA LINC00488 and its effects on thyroid cancer in the view of cell proliferation and apoptosis.
In order to evaluate the effects of LINC00488 on the cellular process of thyroid cancer, we performed a series of in vitro experiments, including cell counting kit-8 (CCK-8) assay, EdU (5-ethynyl-2'-deoxyuridine) assay, flow cytometry, transwell chamber assay, Western blot and RT-qPCR. The target gene of LINC00488 was then identified by bioinformatics analysis (DIANA and TargetScan). Finally, a series of rescue experiments was conducted to validate the effect of LINC00488 and its target genes on proliferation, migration, invasion and apoptosis of thyroid cancer.
Our findings revealed that LINC00488 was highly expressed in thyroid cancer cell lines (BCPAP, BHP5-16, TPC-1 and CGTH-W3) and promoted the proliferation, migration and invasion, while inhibited the apoptosis of thyroid cancer cells (BCPAP and TPC-1). The results of bioinformatics analysis and dual luciferase reporter gene assay showed that LINC00488 could directly bind to miR-376a-3p and down-regulated the expression level of miR-376a-3p. In addition, Paraoxonase-2 (PON2) was a target gene of miR-376a-3p and negatively regulated by miR-376a-3p. Rescue experiment indicated that LINC00488 might enhance PON2 expression by sponging miR-376a-3p in thyroid cancer.
Taken together, our study revealed that lncRNA LINC00488 acted as an oncogenic gene in the progression of thyroid cancer via regulating miR-376a-3p/PON2 axis, which indicated that LINC00488-miR-376a-3p-PON2 axis could serve as novel biomarkers or potential targets for the treatment of thyroid cancer.
长链非编码 RNA(lncRNA)最近被鉴定为多种恶性肿瘤中有影响力的指标。本研究旨在从细胞增殖和凋亡的角度确定一种功能性 lncRNA LINC00488 及其对甲状腺癌的影响。
为了评估 LINC00488 对甲状腺癌细胞过程的影响,我们进行了一系列体外实验,包括细胞计数试剂盒-8(CCK-8)测定、EdU(5-乙炔基-2'-脱氧尿苷)测定、流式细胞术、Transwell 室测定、Western blot 和 RT-qPCR。然后通过生物信息学分析(DIANA 和 TargetScan)鉴定 LINC00488 的靶基因。最后,进行了一系列挽救实验,以验证 LINC00488 及其靶基因对甲状腺癌细胞增殖、迁移、侵袭和凋亡的影响。
我们的研究结果表明,LINC00488 在甲状腺癌细胞系(BCPAP、BHP5-16、TPC-1 和 CGTH-W3)中高表达,并促进甲状腺癌细胞的增殖、迁移和侵袭,同时抑制甲状腺癌细胞的凋亡(BCPAP 和 TPC-1)。生物信息学分析和双荧光素酶报告基因检测结果表明,LINC00488 可以直接与 miR-376a-3p 结合,并下调 miR-376a-3p 的表达水平。此外,对氧磷酶 2(PON2)是 miR-376a-3p 的靶基因,并受 miR-376a-3p 的负调控。挽救实验表明,LINC00488 可能通过海绵吸附 miR-376a-3p 来增强甲状腺癌中的 PON2 表达。
综上所述,本研究表明,lncRNA LINC00488 通过调节 miR-376a-3p/PON2 轴在甲状腺癌的进展中发挥致癌基因作用,表明 LINC00488-miR-376a-3p-PON2 轴可作为甲状腺癌治疗的新型生物标志物或潜在靶点。