miR-338-3p对甲状腺乳头状癌细胞系TPC-1增殖、侵袭、迁移和上皮-间质转化的调控作用  

Regulation of miR-338-3p on proliferation,invasion,migration and epithelial-mesenchymal transformation of thyroid papillary carcinoma cell line TPC-1

作　　者：李涛[1] 李佳[1] 潘婧 梁霄 于丽娜 LI Tao;LI Jia;PAN Jing;LIANG Xiao;YU Lina(Department of Clinical Laboratory,the Second Hospital of Baoding,Baoding 071000,Hebei,China;Department of Nephrology,Rheumatism and Immunology,the Second Hospital of Baoding,Baoding 071000,Hebei,China;Department of Clinical Laboratory,Shijiazhuang Maternal and Child Health Hospital,Shijiazhuang 050050,Hebei,China)

机构地区：[1]保定市第二医院检验科,河北保定071000 [2]保定市第二医院肾病风湿免疫科,河北保定071000 [3]石家庄市妇幼保健院检验科,河北石家庄050050

出　　处：《检验医学》2022年第6期583-589,共7页Laboratory Medicine

基　　金：保定市科技计划项目(1951ZF037)。

摘　　要：目的探讨微小RNA-338-3p(miR-338-3p)过表达靶向沉默信息调节因子(SIRT)6对人甲状腺乳头状癌细胞系TPC-1增殖、侵袭、迁移和上皮-间质转化(EMT)的影响。方法采用荧光素酶报告实验分析miR-338-3p与SIRT6的靶向关系。构建SIRT6 pcDNA载体过表达SIRT6,将miR-338-3p mimic与pcDNA-SIRT6单独或联合转染至TPC-1细胞,根据转染质粒的不同分为6组:对照组(不作处理)、mimic-NC组(转染mimic-NC)、miR-338-3p mimic组(转染miR-338-3p mimic)、pcDNA-SIRT6组(转染pcDNA-SIRT6)、mimic-NC+pcDNA-SIRT6组(共转染mimic-NC和pcDNA-SIRT6)和miR-338-3p mimic+pcDNA-SIRT6组(共转染miR-338-3p mimic和pcDNA-SIRT6)。分别检测各组TPC-1细胞的克隆形成率、侵袭细胞数、划痕愈合率及上皮型钙黏蛋白(E-cad)、神经型钙黏蛋白(N-cad)和波形蛋白的表达量。结果miR-338-3p的靶基因为SIRT6。与对照组比较,miR-338-3p mimic组克隆形成率、侵袭细胞数和划痕愈合率显著降低(P<0.05),E-cad蛋白表达显著升高(P<0.05),N-cad、波形蛋白表达显著降低(P<0.05);pcDNA+SIRT6组克隆形成率、侵袭细胞数和划痕愈合率显著升高,E-cad蛋白表达水平显著降低(P<0.05),N-cad、波形蛋白表达显著升高(P<0.05)。与pcDNA-SIRT6组比较,miR-338-3p mimic+pcDNA-SIRT6组克隆形成率、侵袭细胞数、划痕愈合率显著降低(P<0.05),E-cad蛋白表达显著升高(P<0.05),N-cad、波形蛋白表达显著降低(P<0.05)。结论miR-338-3p过表达靶向SIRT6可抑制TPC-1细胞的增殖、侵袭、迁移和EMT。Objective To investigate the effect of microRNA(miR)-338-3p overexpression of sirtuin(SIRT)6 on proliferation,invasion,migration and epithelial-mesenchymal transformation(EMT)of human thyroid papillary carcinoma cell line TPC-1.Methods Luciferase reporting assay was used to detect the targeting relationship between miR-338-3p and SIRT6.SIRT6 pcDNA vector was constructed to overexpress SIRT6,and miR-338-3p mimic and pcDNA-SIRT6 were transfected into TPC-1 cells either alone or in combination.The transfected plasmids were classified into 6 groups,including control group(no treatment),mimic-NC group(transfected mimic-NC)and miR-338-3p mimic group(transfected miR-338-3p mimic),pcDNA-SIRT6 group(transfected pcDNA-SIRT6),mimic-NC+pcDNA-SIRT6 group(co-transfected mimic-NC and pcDNA-SIRT6)and miR-338-3p mimic+pcDNA-SIRT6 group(co-transfected with miR-338-3p mimic and pcDNA-SIRT6).The clone genesis rate,invasive cell number,scratch healing rate of TPC-1 cells and the expression levels of E-cadherin(E-cad),N-cadherin(N-cad)and Vimentin in epithelial cells were determined.Results The target gene of miR-338-3p was SIRT6.Compared with the control group,clone genesis rate,invasive cell number and scratch healing rate in miR-338-3p mimic group were decreased(P<0.05).The protein expression of E-cad was increased(P<0.05),while the protein expressions of N-cad and Vimentin were decreased(P<0.05).The clone genesis rate,invasive cell number and scratch healing rate were increased in pcDNA+SIRT6 group.The protein expression level of E-cad was decreased(P<0.05),while the protein expressions of N-cad and Vimentin were increased(P<0.05).Compared with pcDNA-SIRT6 group,clone genesis rate,invasive cell number and scratch healing rate in miR-338-3p mimic+pcDNA-SIRT6 group were decreased(P<0.05),while E-cad protein expression was increased(P<0.05).The protein expressions of N-cad and Vimentin were decreased(P<0.05).Conclusions The miR-338-3p overexpression targets SIRT6 to inhibit TPC-1 cell proliferation,invasion,migration and EMT.

关 键 词：微小RNA-338-3p 沉默信息调节因子6 甲状腺乳头状癌 

分 类 号：R446.7[医药卫生—诊断学]