实用医学杂志 ›› 2022, Vol. 38 ›› Issue (5): 565-570.doi: 10.3969/j.issn.1006⁃5725.2022.05.009

• 基础研究 • 上一篇    下一篇

microRNA⁃7⁃5p在甲状腺乳头状癌中的表达及机制

秦小静1,2,3 范会利1,3 林旭1,3 王冬梅1,3 薛刚1,3 吴靖芳1,3   

  1. 1 河北北方学院形态学实验室(河北张家口 075000);2 河北中医学院(石家庄 050091);3 张家口市重点实 验室(河北张家口 075000)

  • 出版日期:2022-03-10 发布日期:2022-03-10
  • 通讯作者: 吴靖芳 E⁃mail:wjfxg@ 163.com;薛刚 E⁃mail:109568894@qq.com
  • 基金资助:
    河北省研究生创新基金(编号:CXZZSS20211131);张家口市科技计划项目(编号:1821030D)

Expression and mechanism of microRNA⁃7⁃5p in papillary thyroid carcinoma 

QIN Xiaojing*,FAN Huili, LIN Xu,WANG Dongmei,XUE Gang,WU Jingfang.    

  1. Laboratory of Morphology,Hebei North University,Zhangji⁃ akou 075000,China;*Hebei University of Chinese Medicine,Shijiazhuang 050091,China;*Key Laboratory of Zhangjiakou,Zhangjiakou 075000,China 

  • Online:2022-03-10 Published:2022-03-10
  • Contact: WU Jingfang E⁃mail:wjfxg@ 163.com;XUE Gang E⁃mail:109568894@qq.com

摘要:

目的 探讨 microRNA⁃7⁃5p(miR⁃7⁃5p)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC 组织和细胞系的表达以及对 PTC 细胞增殖、侵袭及迁移能力的影响及相关分子机制。方法 实时荧光定 量聚合酶链反应(quantitative real⁃time PCR,qRT⁃PCR)检测 52 PTC 及癌旁组织和人 PTC 细胞系 TPC⁃1 BCPAP、K1及人正常甲状腺滤泡细胞系Nthy⁃ori 3⁃1中miR⁃7⁃5p的表达情况。分别将miR⁃7⁃5p mimics/inhibi⁃ tor、mimics⁃NC/NC⁃inhibitor(NC⁃in)以脂质体 LipofectaminTM 3000 转染至 TPC⁃1 K1 细胞分为 3 组:(1)空白 对照组(TPC⁃1/K1 组);(2)阴性对照组(mimics⁃NC 组/NC⁃in 组);(3)实验组(mimics⁃7⁃5p 组/inhibitor⁃7⁃5p 组)。以生长曲线、小室侵袭实验和划痕实验检测 3 组细胞增殖、侵袭和迁移能力。双荧光素酶报告基因 实验检测 miR⁃7⁃5p 与三叶因子 3(TFF3)3′UTR 区结合情况,Western blot 检测 miR⁃7⁃5p TFF3 蛋白表达 的影响。结果 PTC 组织和 3 PTC 细胞系中 miR⁃7⁃5p 表达水平明显低于癌旁组织和 Nthy⁃ori 3⁃1 正常滤 泡上皮细胞(P < 0.01),且不同临床分期、肿瘤大小和淋巴结转移的 PTC 患者 miR⁃7⁃5p 表达差异有统计学 意义(P < 0.01)。上调 miR⁃7⁃5p 后,实验组 TPC⁃1 细胞的增殖活性、侵袭和迁移能力明显下降(P < 0.05)。 双荧光素酶报告实验结果表明 mimics⁃7⁃5p/inhibitor⁃7⁃5p 组的荧光酶活性低于/高于对照组(P < 0.01)。 结论 miR⁃7⁃5p在PTC组织中低表达,miR⁃7⁃5p可以通过靶向TFF3抑制PTC细胞的增殖、侵袭及迁移能力。

关键词:

miR?7?5p, 甲状腺乳头状癌, 三叶因子3

Abstract:

Objective To explore the expression of microRNA ⁃7⁃5p(miR ⁃7⁃5p)in papillary thyroid carcinoma(PTC)tissues,para⁃carcinoma,as well as cell lines and its influenceon the proliferation,invasion and migration of PTC cells and its related molecular mechanism. Methods Quantitative Real⁃Time PCR(qRT⁃PCR was used to detect the expression of miR⁃7⁃5p in 52 cases of PTC,para⁃carcinoma and human PTC cell lines TPC⁃1 BCPAP,K1 and human normal thyroid follicular cell line Nthy⁃ori 3⁃1,respectively. MiR⁃7⁃5p mimics and mimics ⁃NC were transfected into TPC⁃1 cells and K1cells with LipofectaminTM 3000 and divided into blank control group,negative control group(mimics⁃NC/NC⁃in group)and experimental group(mimics⁃7⁃5p/inhibitor⁃7⁃5p group). The proliferation,invasion and migration abilities of cells in 3 groups were tested by growth curve,transwell invasion and scratch. The binding of miR ⁃7⁃5p to the TFF3⁃3′UTR was detected by dual luciferase reporter gene experi⁃ ment,and effect of miR⁃7⁃5p on TFF3 expression by western blot. Results The expression of miR⁃7⁃5p in PTC and the 3 PTC cell lines was significantly lower than that in adjacent tissues and Nthy⁃ori 3⁃1 cell(P < 0.01). Mir⁃ 7⁃5p expression was significantly different in PTC patients with different clinical stages,tumor size and lymph node metastasis(P < 0.01). After the up⁃regulation of miR⁃7⁃5p,the proliferation,invasion and migration abilities of TPC⁃1 and K1cells decreased significantly(P < 0.05). The double luciferase reporter gene experiment showed that the luciferase activity value in mimics⁃7⁃5p/inhibitor⁃7⁃5p group was lower/or higher when compared that in the control group(P < 0.01). Conclusion The expression of miR⁃7⁃5p is low in PTC tissues,and miR⁃7⁃5p can inhibit the proliferation,invasion and migration of PTC cells by targeting TFF3.

Key words:

miR?7?5p, papillary thyroid carcinoma, trefoil factor 3(TFF3)