Zhou Qinyi, Chen Jun, Feng Jialin, Wang Jiadong
Department of Head and Neck Surgery, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, #145 Shandong Road, Huangpu District, Shanghai, 200001, China.
Tumour Biol. 2016 Mar;37(3):3105-13. doi: 10.1007/s13277-015-4149-9. Epub 2015 Oct 1.
The purposes of this study were to investigate the potential roles of long noncoding RNA (lncRNA) PVT1 in thyroid cancer cell proliferation and to explore their possible mechanisms. A total of 84 patients who were diagnosed as having thyroid cancer (papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), and anaplastic thyroid carcinoma (ATC)) in Renji Hospital were enrolled in this study. Expressions of lncRNA PVT1 in thyroid cancer tissues and cell lines (IHH-4, FTC-133, and 8505C) were analyzed using RT-polymerase chain reaction (PCR) and western blotting analysis. The effects of lncRNA PVT1 expression on thyroid cancer cell proliferation and cell cycle were analyzed using flow cytometry. Furthermore, the effects of lncRNA expression on thyroid-stimulating hormone receptor (TSHR) expression and polycomb enhancer of zeste homolog 2 (EZH2) were also analyzed using RNA immunoprecipitation (RIP) assay and chromatin immunoprecipitation (ChIP) assay, respectively. Compared to the controls, lncRNA PVT1 was significantly up-regulated in thyroid tissues, as well as in three kinds of tumor cell lines (P < 0.05). Silenced PVT1 significantly inhibited thyroid cell line IHH-4, FTC-133, and 8505C cell proliferation and arrested cell cycle at G0/G1 stage and significantly decreased cyclin D1 and TSHR expressions (P < 0.05). Moreover, lncRNA PVT1 could be enriched by EZH2, and silencing PVT1 resulted in the decreased recruitment of EZH2. This study suggested that lncRNA PVT1 may contribute to tumorigenesis of thyroid cancer through recruiting EZH2 and regulating TSHR expression.
本研究旨在探讨长链非编码RNA（lncRNA）PVT1在甲状腺癌细胞增殖中的潜在作用，并探索其可能的机制。本研究纳入了84例在仁济医院被诊断为患有甲状腺癌（乳头状甲状腺癌（PTC）、滤泡状甲状腺癌（FTC）和未分化甲状腺癌（ATC））的患者。采用逆转录聚合酶链反应（PCR）和蛋白质印迹分析技术分析lncRNA PVT1在甲状腺癌组织和细胞系（IHH-4、FTC-133和8505C）中的表达。采用流式细胞术分析lncRNA PVT1表达对甲状腺癌细胞增殖和细胞周期的影响。此外，还分别采用RNA免疫沉淀（RIP）试验和染色质免疫沉淀（ChIP）试验分析lncRNA表达对促甲状腺激素受体（TSHR）表达和zeste同源物2多梳增强子（EZH2）的影响。与对照组相比，lncRNA PVT1在甲状腺组织以及三种肿瘤细胞系中均显著上调（P<0.05）。沉默PVT1可显著抑制甲状腺细胞系IHH-4、FTC-133和8505C细胞的增殖，并使细胞周期停滞在G0/G1期，同时显著降低细胞周期蛋白D1和TSHR的表达（P<0.05）。此外，lncRNA PVT1可被EZH2富集，沉默PVT1可导致EZH2的募集减少。本研究表明，lncRNA PVT1可能通过募集EZH2和调节TSHR表达促进甲状腺癌的发生。
