a Department of Nuclear Medicine , Yunnan Cancer Hospital & The Third Affiliated Hospital of Kunming Medical University & Yunnan Cancer Center , Kunming , Yunnan , China.
Cell Cycle. 2019 Jan;18(2):167-203. doi: 10.1080/15384101.2018.1560203. Epub 2018 Dec 30.
Considering the resistance of papillary thyroid cancer (PTC) I therapy, this study was designed to find a solution at molecular respect. By probing into lncRNA-NEAT1/miR-101-3p/FN1 axis and PI3K/AKT signaling pathway, this study provided a potential target for PTC therapy. I-resistant cell lines were established by continuous treatment with median-lethal I. Bioinformatic analysis was applied to filtrate possible lncRNA/miRNA/mRNA and related signaling pathway. Luciferase reporter assay was employed in the verification of the targeting relationship between lncRNA and miRNA as well as miRNA and mRNA. MTT assay and flow cytometry assay were performed to observe the impact of NEAT1/miR-101-3p/FN1 on cell viability and apoptosis in radioactivity iodine (RAI)-resistant PTC cell lines, respectively. Western blot and qRT-PCR were conducted to measure the expression of proteins and mRNAs in RAI-resistant PTC tissues and cells. Meanwhile, endogenous PTC mice model were constructed, in order to verify the relation between NEAT1 and RAI-resistance in vivo. NEAT1 was over-expressed in RAI-resistant PTC tissues and cell lines and could resist RAI by accelerating proliferation accompanied by suppressing apoptosis. It indicated that overexpressed NEAT1 restrained the damage of RAI to tumor in both macroscopic and microcosmic. Besides, NEAT1/miR-101-3p exhibited a negative correlation by directly targeting each other. The expression of FN1, an overexpressed downstream protein in RAI-resistance PTC tissues, could be tuned down by miR-101-3p, while the decrease could be restored by NEAT1. In conclusion, both in vitro and in vivo, NEAT1 suppression could inhibit I resistance of PTC by upregulating miR-101-3p/FN1 expression and inactivated PI3K/AKT signaling pathway both in vitro and in vivo.
考虑到甲状腺乳头状癌 (PTC) 的放射性碘 (RAI) 治疗抵抗性，本研究旨在从分子水平寻找解决方案。通过探讨 lncRNA-NEAT1/miR-101-3p/FN1 轴和 PI3K/AKT 信号通路，为 PTC 的治疗提供了潜在的靶点。通过连续用中等致死剂量的 I 处理来建立耐 I 细胞系。应用生物信息学分析筛选可能的 lncRNA/miRNA/mRNA 及其相关信号通路。荧光素酶报告基因检测用于验证 lncRNA 与 miRNA 以及 miRNA 与 mRNA 之间的靶向关系。MTT 检测和流式细胞术检测分别用于观察 NEAT1/miR-101-3p/FN1 对放射性碘耐药 PTC 细胞系细胞活力和凋亡的影响。Western blot 和 qRT-PCR 用于检测放射性碘耐药 PTC 组织和细胞中蛋白质和 mRNAs 的表达。同时，构建内源性 PTC 小鼠模型，以验证 NEAT1 在体内与放射性碘耐药的关系。NEAT1 在放射性碘耐药的 PTC 组织和细胞系中高表达，通过加速增殖同时抑制凋亡来抵抗放射性碘。结果表明，过表达的 NEAT1 从宏观和微观上抑制了放射性碘对肿瘤的损伤。此外，NEAT1/miR-101-3p 通过直接靶向对方表现出负相关。FN1 是放射性碘耐药 PTC 组织中过表达的下游蛋白，miR-101-3p 可以下调其表达，而 NEAT1 可以恢复其表达。总之，在体外和体内，通过上调 miR-101-3p/FN1 表达和抑制 PI3K/AKT 信号通路，抑制 NEAT1 可以抑制 PTC 的 I 耐药性。
