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例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Lipofectamin^(TM) 3000转染至TPC⁃1和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对TFF3蛋白表达的影响。结果PTC组织和3株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。
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 Lipofectamin^(TM)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 experiment,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.
作者
秦小静
范会利
林旭
王冬梅
薛刚
吴靖芳
QIN Xiaojing;FAN Huili;LIN Xu;WANG Dongmei;XUE Gang;WU Jingfang(Laboratory of Morphology,Hebei North University,Zhangjiakou 075000,China;Hebei University of Chinese Medicine,Shijiazhuang 050091,China;Key Laboratory of Zhangjiakou,Zhangjiakou 075000,China;不详)
出处
《实用医学杂志》
CAS
北大核心
2022年第5期565-570,共6页
The Journal of Practical Medicine
基金
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)。
