Tamagawa Shunji, Enomoto Keisuke, Gunduz Esra, Gunduz Mehmet, Sato Fuyuki, Uchino Shinya, Muragaki Yasuteru, Hotomi Muneki
Department of Otolaryngology-Head and Neck Surgery, Wakayama Medical University, Wakayama, Wakayama 641-8509, Japan.
Department of Pathology, Wakayama Medical University, Wakayama, Wakayama 641-8509, Japan.
Oncol Lett. 2020 Oct;20(4):3. doi: 10.3892/ol.2020.11864. Epub 2020 Jul 14.
Anaplastic thyroid cancer (ATC) remains a cancer with one of the worst prognoses, despite novel targeted therapies. The median survival rate has not improved for decades. Epithelial-to-mesenchymal transition (EMT) is a crucial step in physiological processes and in cancer progression, but the underlying mechanisms are not yet fully understood. The current study examined the role of microRNA (miR)-200b in mesenchymal-to-epithelial transition in ATC. Total RNA and miR isolation were performed from ATC cell lines transfected with a miR-200b mimic. After miR-200b mimic transfection, expression levels of E-cadherin, vimentin and zinc finger E-box binding homeobox 1 (ZEB1) were confirmed by reverse transcription-quantitative PCR and western blotting. Additionally, cell migration was evaluated using miR-200b mimic and scrambled negative control-transfected cells. A total of 14 human ATC and 15 non-cancerous human thyroid tissues were immunohistochemically stained and scored as controls for E-cadherin, vimentin and ZEB1. In ATC tissues and cell lines, the mesenchymal marker ZEB1 was significantly upregulated and the epithelial marker E-cadherin was significantly downregulated. Additionally, the mesenchymal marker vimentin was significantly upregulated in ATC tissues and in one ATC cell line. MiR-200b mimic transfection significantly increased vimentin and ZEB1 expression, but E-cadherin expression remained below the measurement sensitivity. Furthermore, miR-200b overexpression decreased cell migration. The current study suggested that miR-200b may regulate the expression levels of mesenchymal markers such as vimentin and ZEB1 in ATC and may promote mesenchymal-to-epithelial transition.
尽管有新型靶向疗法,但间变性甲状腺癌(ATC)仍然是预后最差的癌症之一。几十年来,其平均生存率并未提高。上皮-间质转化(EMT)是生理过程和癌症进展中的关键步骤,但其潜在机制尚未完全明确。本研究探讨了微小RNA(miR)-200b在ATC间质-上皮转化中的作用。从转染了miR-200b模拟物的ATC细胞系中提取总RNA和miR。转染miR-200b模拟物后,通过逆转录定量PCR和蛋白质印迹法确认E-钙黏蛋白、波形蛋白和锌指E盒结合同源框1(ZEB1)的表达水平。此外,使用转染了miR-200b模拟物和乱序阴性对照的细胞评估细胞迁移。对14例人ATC组织和15例非癌性人甲状腺组织进行免疫组织化学染色,并对E-钙黏蛋白、波形蛋白和ZEB1进行评分作为对照。在ATC组织和细胞系中,间质标志物ZEB1显著上调,上皮标志物E-钙黏蛋白显著下调。此外,间质标志物波形蛋白在ATC组织和一种ATC细胞系中显著上调。转染miR-200b模拟物显著增加了波形蛋白和ZEB1的表达,但E-钙黏蛋白的表达仍低于测量灵敏度。此外,miR-200b过表达减少了细胞迁移。本研究表明,miR-200b可能调节ATC中间质标志物如波形蛋白和ZEB1的表达水平,并可能促进间质-上皮转化。