miR-375-3p过表达的甲状腺乳头状癌细胞增殖和侵袭能力变化  

Effects of over-expression of miR-375-3p on proliferation and invasion of thyroid papillary carcinoma cells

作　　者：卢金喜 余红梅 齐孝安 方超 魏新宝 李立鑫 LU Jinxi;YU Hongmei;QI Xiaoan;FANG Chao;WEI Xinbao;LI Lixin(Department of General Surgery,Xinzhou District People's Hospital,Wuhan 430400,China)

机构地区：[1]武汉市新洲区人民医院普外科,武汉430400

出　　处：《山东医药》2023年第14期50-54,共5页Shandong Medical Journal

摘　　要：目的探讨miR-375-3p过表达后甲状腺乳头状癌细胞增殖和侵袭能力以及PI3K/AKT/EMT信号通路相关蛋白的变化。方法取人甲状腺上皮细胞系Nthy-ori3-1和人甲状腺乳头状癌细胞系TPC-1、BHP5-16、BHP2-7、K-1,采用RT-qPCR法检测细胞miR-375-3p表达。以四种甲状腺乳头状癌细胞中miR-375-3p表达最低的细胞为研究对象,将其分为转染组和阴性对照组,转染组通过转染miR-375-3p模拟物miR-375-3p-mimics上调细胞中miR-375-3p表达,阴性对照组转染阴性对照序列scramble。采用MTT法检测两组细胞增殖能力,Transwell实验观察细胞侵袭能力,Western blotting法检测细胞PI3K/AKT信号通路相关蛋白p-PI3K、p-AKT和上皮间质转化(EMT)相关蛋白E钙黏蛋白(E-cadherin)、N钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达。取甲状腺乳头状癌细胞,随机分为阴性对照组、miR-375-3p过表达组和miR-375-3p过表达+IGF-1组,miR-375-3p过表达组、阴性对照组分别转染miR-375-3p模拟物miR-375-3p mimics和阴性对照序列scramble,miR-375-3p过表达+IGF-1组转染miR-375-3p mimics并加入PI3K/AKT信号通路激动剂胰岛素样生长因子1(IGF-1)。观察三组细胞增殖和侵袭能力。结果人甲状腺乳头状癌细胞TPC-1、BHP5-16、BHP2-7、K-1中miR-375-3p表达水平均低于人甲状腺上皮细胞Nthy-ori3-1(P均<0.05)。miR-375-3p过表达组转染24、48、72 h的OD值低于阴性对照组,侵袭细胞数少于阴性对照组(P<0.05或<0.01)。与阴性对照组比较,miR-375-3p过表达组p-PI3K和p-AKT表达水平降低,E-cadherin表达水平升高,N-cadherin、波形蛋白表达水平降低(P均<0.05)。加入IGF-1后,与miR-375-3p过表达组比,miR-375-3p过表达+IGF-1组细胞增殖OD值和细胞侵袭数均增加(P均<0.05)。结论miR-375-3p在甲状腺乳头状癌细胞系中下调表达,miR-375-3p过表达可抑制细胞增殖、侵袭过程,其机制可能与抑制PI3K/AKT/EMT信号通路有关。Objective To investigate the effects of over-expression of miR-375-3p on the proliferation and invasion of thyroid papillary carcinoma cells,and to clarify the mechanism.Methods The miR-375-3p expression in the human thyroid epithelial cell line Nthy-ori3-1 and human thyroid papillary carcinoma cell lines TPC-1,BHP5-16,BHP2-7,and K-1 was detected by RT-qPCR.Thyroid papillary carcinoma cell line TPC-1 which had the lowest expression of miR-375-3p was taken and divided into the transfected group and negative control group.In the transfected group,we up-regulated the expression of miR-375-3p in thyroid cancer cell line TPC-1 by transfecting miR-375-3p mimics,while cells in the the negative control group were transfected with negative control sequences(scramble).The cell proliferation capacities of two groups were observed by MTT assay.Invasion capacities were observed by Transwell assay.The expression levels of PI3K/AKT signaling pathway-related proteins p-PI3K and p-AKT and epithelial-mesenchymal transition(EMT)-related proteins E-cadherin,N-cadherin,and Vimentin were detected by Western blotting.PTC cells were randomly divided into the nega⁃tive control group,miR-375-3p over-expression group,and miR-375-3p over-expression+IGF-1 group.Cells in the miR-375-3p over-expression group and negative control group were transfected with miR-375-3p mimics(miR-375-3p mimics)and negative control sequence(scramble)through LifefectamineTM3000,respectively.Cells in the miR-375-3p over-ex⁃pression+IGF-1 group were treated with miR-375-3p mimics and the PI3K/AKT signal pathway agonist insulin-like growth factor-1(IGF-1).The proliferative and invasive capacities of cells in the three groups were observed.Results The ex⁃pression levels of miR-375-3p in cell lines TPC-1,BHP5-16,BHP2-7 and K-1 were significantly lower than that of normal human thyroid epithelial cell line Nthy-ori3-1(all P<0.05).The OD values of the miR-375-3p over-expression group were significantly lower than those of the negative control group at 24,48 and 72

关 键 词：miR-375-3p 细胞增殖 细胞侵袭 上皮间质转化 PI3K/AKT信号通路 甲状腺癌细胞 甲状腺癌 

分 类 号：R736.1[医药卫生—肿瘤]