Abstract:

Objective　To investigate the role of long noncoding RNA (lncRNA) LY6E-DT in the proliferation and migration of thyroid cancer cells regulating miR-148a expression. Methods　The research was from March 2023 to October 2024. The expression level of LY6E-DT in thyroid cancer tissues was analyzed using the UCSC Xena database. The real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression levels of LY6E-DT in the thyroid cancer cell lines (FTC-133, TPC-1, KTC-1, and 8505C) and thyroid epithelial cells HTori-3. The TPC-1 cells were divided into an si-LY6E-DT group and an si-NC group according to the transfection methods. The clone formation ability of the TPC-1 cells was detected by the clone formation assay. The migration ability of the TPC-1 cells was detected by the Transwell assay. The interaction between LY6E-DT and miR-148a was verified by the dual luciferase reporter assay and qPCR. The expression levels of p-PI3K, p-mTOR, p27, and p-GLUT4 proteins in the PI3K/AKT signaling pathway in the TPC-1 cells were detected by the Western blot. The data were compared between two groups by the t test and between multiple groups by one-way analysis of variance. Results　The expression level of LY6E-DT in the thyroid cancer tissues was significantly higher than that in the adjacent tissues (P<0.01). The expression levels of LY6E-DT in the FTC-133, TPC-1, KTC-1, and 8505C cells were significantly higher than that in the HTori-3 cells (all P<0.01). The expression level of LY6E-DT in the si-LY6E-DT group was lower than that in the si-NC group (1.06±0.59 vs. 7.11±1.53; t=7.37, P<0.01). The numbers of clones formed in the si-LY6E-DT group and the si-NC group were 38.01±9.63 and 113.10±13.33 (t=4.60, P<0.01), and the numbers of cell migration were 45.24±5.99 and 85.65±5.43 (t=4.99, P<0.01). LY6E-DT could target and negatively regulate the expression of miR-148a (P<0.01). Compared with those in the si-NC group, the expression levels of p-PI3K, p-mTOR, p27, and p-GLUT4 proteins in the PI3K/AKT signaling pathway in the TPC-1 cells in the si-LY6E-DT group were lower (all P<0.01). Conclusions　LY6E-DT is over-expressed in thyroid cancer tissues and cell lines. Knockdown of LY6E-DT can inhibit the proliferation and migration of thyroid cancer TPC-1 cells by targeted regulation of miR-148a expression.

Key words:

Thyroid cancer, Long noncoding RNA, LY6E-DT, MiR-148a, Proliferation, Migration

摘要：

目的　探讨长链非编码RNA（lncRNA）LY6E-DT通过调控miR-148a表达在甲状腺癌细胞增殖和迁移中的作用。方法　研究时间为2023年3月至2024年10月。用UCSC Xena数据库分析甲状腺癌组织中LY6E-DT的表达水平。实时定量聚合酶链式反应（qPCR）检测甲状腺癌细胞系（FTC-133、TPC-1、KTC-1、8505C）和甲状腺上皮细胞HTori-3中LY6E-DT的表达水平。根据转染方法将TPC-1细胞分为si-LY6E-DT组和si-NC组。克隆形成实验检测TPC-1细胞的克隆形成能力。Transwell实验检测TPC-1细胞的迁移能力。双荧光素酶报告实验与qPCR验证LY6E-DT与miR-148a的相互作用。Western blot检测TPC-1细胞中PI3K/AKT信号通路蛋白p-PI3K、p-mTOR、p27、p-GLUT4的表达水平。两组间比较采用t检验，多组间比较采用单因素方差分析。结果　甲状腺癌组织中LY6E-DT表达水平高于癌旁组织（P<0.01）。FTC-133、TPC-1、KTC-1、8505C细胞中LY6E-DT表达水平均高于HTori-3细胞（均P<0.01）。si-LY6E-DT组和si-NC组TPC-1细胞中LY6E-DT表达水平分别为1.06±0.59和7.11±1.53（t=7.37，P<0.01）。si-LY6E-DT组和si-NC组克隆形成数分别为（38.01±9.63）个和（113.10±13.33）个（t=4.60，P<0.01），细胞迁移数分别为（45.24±5.99）个和（85.65±5.43）个（t=4.99，P<0.01）。LY6E-DT能够靶向并负调控miR-148a的表达（P<0.01）。与si-NC组比较，si-LY6E-DT组TPC-1细胞中PI3K/AKT信号通路p-PI3K、p-mTOR、p27、p-GLUT4蛋白表达水平均较低（均P<0.01）。结论　LY6E-DT在甲状腺癌组织和细胞系中均表达升高。敲降LY6E-DT能通过靶向调控miR-148a表达抑制甲状腺癌TPC-1细胞的增殖和迁移。

关键词:

甲状腺癌, 长链非编码RNA, LY6E-DT, miR-148a, 增殖, 迁移