标题：miR-214-5p靶向FGFRL1对甲状腺癌细胞TPC-1增殖、侵袭和迁移影响

      作者：高建华 1，杜甲录 1，李今 2    高建华 1，杜甲录 1，李今 21.榆林市第一医院普通外科二病区，陕西 榆林 719000；2.西北妇女儿童医院乳腺甲状腺外科，陕西 西安 710061

      卷次： 2023年34卷16期

      【摘要】 目的 探讨微小RNA-214-5p (miR-214-5p)靶向成纤维细胞生长因子受体样 1 (FGFRL1)调控甲状腺癌细胞增殖、侵袭和迁移的机制。方法 2019年1~5月收集来自榆林市第一医院的47例甲状腺癌患者的癌组织及其癌旁组织，体外培养甲状腺癌BCPAP、TPC-1、SW1736细胞和正常甲状腺HT-ori3细胞，采用qRT-PCR法检测miR-214-5p与 FGFRL1的表达；免疫组化检测甲状腺癌组织中 FGFRL1阳性率；miR-214-5p模拟物/抑制剂(miR-214-5p mimics/anti-miR-214-5p)及阴性对照(miR-con/anti-miR-con)、FGFRL1小干扰RNA (si-FGFRL1)及其阴性对照(si-con)、miR-214-5p mimics与 pcDNA-con、miR-214-5p mimics与 FGFRL1过表达载体(pcDNA-FGFRL1)转染至 TPC-1细胞；采用甲基噻唑基四唑(MTT)与 Transwell迁移及侵袭实验分别检测miR-214-5p、FGFRL1对 TPC-1细胞增殖、迁移及侵袭能力的影响。双荧光素酶报告基因实验验证miR-214-5p与 FGFRL1的关系。结果 与癌旁组织比较，甲状腺癌组织中miR-214-5p的表达量降低，FGFRL1的表达量升高，FGFRL1阳性率升高，差异均有统计学意义(P<0.05)；甲状腺癌细胞中miR-214-5p表达水平低于正常甲状腺细胞，而FGFRL1表达水平高于正常甲状腺细胞，差异均有统计学意义(P<0.05)；转染miR-214-5p mimics后，TPC-1细胞存活率降低，迁移及侵袭细胞数减少，差异均有统计学意义(P<0.05)；转染 anti-miR-214-5p后，TPC-1细胞存活率升高，迁移及侵袭细胞数增多，差异均有统计学意义(P<0.05)；敲减FGFRL1可明显抑制TPC-1细胞增殖、迁移及侵袭，差异均有统计学意义(P<0.05)；双荧光素酶报告基因实验证明miR-214-5可靶向结合 FGFRL1；FGFRL1过表达可逆转上调miR-214-5p表达对TPC-1细胞增殖、迁移及侵袭的抑制作用。结论 miR-214-5p通过靶向FGFRL1而减弱甲状腺癌细胞增殖、迁移及侵袭能力。
      【关键词】 甲状腺癌；微小RNA-214-5p；成纤维细胞生长因子受体样1；增殖；迁移；侵袭
      【中图分类号】 R736.1 【文献标识码】 A 【文章编号】 1003—6350（2023）16—2281—06

Mechanism of miR-214-5p targeting FGFRL1 in regulating proliferation, invasion, and migration of thyroidcancer cells TPC-1.
GAO Jian-hua 1, DU Jia-lu 1, LI Jin 2. 1. The Second Ward, Department of General Surgery, YulinFirst Hospital, Yulin 719000, Shaanxi, CHINA; 2. Department of Breast and Thyroid Surgery, Northwest Women's andChildren's Hospital, Xi'an 710061, Shaanxi, CHINA
【Abstract】 Objective To investigate the mechanism by which microRNA-214-5p (miR-214-5p) targets fibro-blast growth factor receptor-like 1 (FGFRL1) to regulate proliferation, invasion, and migration of thyroid cancer cells.Methods Cancer tissues and adjacent tissues of 47 thyroid cancer patients from Yulin First Hospital were collectedfrom January 2019 to May 2019, and thyroid cancer BCPAP, TPC-1, SW1736 cells and normal thyroid HT-ori3 cellswere cultured in vitro. qRT-PCR was used to detect the expression of miR-214-5p and FGFRL1. The positive rate of FG-FRL1 in thyroid cancer tissue was detected by immunohistochemistry. miR-con, miR-214-5p mimics, anti-miR-con, an-ti-miR-214-5p, si-con, si-FGFRL1, miR-214-5p mimics and pcDNA-con, miR-214-5p mimics, and pcDNA-FGFRL1were transfected into TPC-1 cells. MTT was used to detect the effect of miR-214-5p and FGFRL1 on the proliferationof TPC-1 cells. Transwell migration and invasion assays were performed to examine the effects of miR-214-5p and FG-FRL1 on the migration and invasion of TPC-1 cells. The dual luciferase reporter gene assay was used to verify the rela-tionship between miR-214-5p and FGFRL1. Results Compared with adjacent tissues, the expression of miR-214-5pin thyroid cancer tissues was decreased, the expression of FGFRL1 was increased, and the positive rate of FGFRL1 wasincreased, all with statistically significant differences (P<0.05). The expression level of miR-214-5p in thyroid cancercells was lower than that in normal thyroid cells, while the expression level of FGFRL1 was higher than that in normalthyroid cells, with statistically significant differences (P<0.05). After transfection with miR-214-5p mimics, the survivalrate of TPC-1 cells, migration and invasion cells were significantly reduced (P<0.05). After transfection with an-ti-miR-214-5p, the survival rate of TPC-1 cells increased, and the number of migratory and invasive cells increased,with statistically significant differences (P<0.05). Knockdown of FGFRL1 significantly inhibited proliferation, migra-tion, and invasion of TPC-1 cells (P<0.05). The dual luciferase reporter assay demonstrated that miR-214-5 can targetFGFRL1. Overexpression of FGFRL1 reversed the inhibition of proliferation, migration, and invasion of TPC-1 cells byupregulating miR-214-5p expression. Conclusion miR-214-5p attenuates the proliferation, migration, and invasionof thyroid cancer cell by targeting FGFRL1.
      【Key words】 Thyroid cancer; miR-214-5p; FGFRL1; Proliferation; Migration; Invasion