摘要
目的探讨MAPT-IT1在乳腺癌中表达的临床意义及其体外生物学效应。方法TCGA数据库分析MAPT-IT1在乳腺癌中的表达,收集64例乳腺癌及正常癌旁组织,培养人乳腺癌MDA-MB-231及MCF-7细胞,通过细胞转染技术过表达MAPT-IT1,回复表达miR-181a-5p,将MDA-MB-231细胞分为:空白A组、过表达A组及恢复A组;将MCF-7细胞分为:空白B组、过表达B组及回复B组。实时荧光定量PCR(RT-qPCR)实验检测组织或各组细胞MAPT-IT1及其预测靶基因miR-181a-5p及MAPT mRNA表达水平,Western blot检测MAPT蛋白表达水平,CCK-8实验检测细胞增殖能力的变化,Transwell侵袭实验检测细胞侵袭能力的变化。结果MAPT-IT1在正常癌旁组织及乳腺癌组织中表达分别为0.011±0.002及0.028±0.003;MAPT-IT1在乳腺癌组织中表达显著高于正常癌旁组织(t=4.08,P<0.001),其高表达与患者更早的临床分期、ER阳性及预后时间延长相关(P均<0.05)。空白A组、过表达A组及回复A组MAPT-IT1表达分别为(1.000±0.078)、(8.597±0.320)、(8.540±0.177),miR-181a-5p表达分别为(1.000±0.027)、(0.263±0.024)、(4.433±0.239),MAPT表达分别为(1.000±0.071)、(3.297±0.243)、(0.497±0.029)。空白B组、过表达B组及回复B组MAPT-IT1表达分别为(1.000±0.081)、(5.716±0.309)、(5.288±0.176),miR-181a-5p表达分别为(1.000±0.024)、(0.291±0.022)、(3.648±0.073),MAPT表达分别为(1.000±0.054)、(3.309±0.177)、(0.883±0.075)。过表达MAPT-IT1后,MDA-MB-231及MCF-7细胞miR-181a-5p表达下调(P<0.05),而MAPT的表达水平显著增高(P<0.001),同时细胞增殖及侵袭能力显著降低(P<0.05)。恢复表达miR-181a-5p则可下调MAPT,促进细胞增殖及侵袭能力(P<0.05)。结论MAPT-IT1的过表达可显著下调乳腺癌细胞miR-181a-5p,介导MAPT表达上调及细胞体外恶性表型的抑制。
Objective To investigate the clinical significance of MAPT-IT1 in breast cancer and its biological effect in vitro.Methods The expression of MAPT-IT1 in breast cancer was analyzed by TCGA database.64 cases of breast cancer and normal adjacent tissues were collected.Human breast cancer MDA-MB-231 and MCF-7 cells were cultured and overexpressed MAPT-IT1 or rescue miR-181a-5p by cell transfection.MDA-MB-231 cells were divided into blank group A,overexpression group A and recovery group A;MCF-7 cells were divided into blank group B,overexpression group B and recovery group B.Real time quantitative PCR(RT-qPCR)was used to detect the expression of MAPT-IT1,miR-181a-5p and MAPT mRNA.Western blot was used to detect the expression of MAPT protein.CCK-8 assay was used to detect cell proliferation,and Transwell invasion assay was used to detect cell invasion.Results The expression of MAPT-IT1 in normal paracancerous tissues and breast cancer tissues was 0.011±0.002 and 0.028±0.003 respectively.The expressions of MAPT-IT1 in breast cancer tissues were significantly higher than those in adjacent normal tissues,and high expression of MAPT-IT1 was correlated with early tumor progression,ER positive and prolonged prognosis(P<0.05).In blank group A,overexpression group A and recovery group A,the expressions of MAPT-IT1 were 1.000±0.078,8.597±0.320 and 8.540±0.177,miR-181a-5p were 1.000±0.027,0.263±0.024,4.433±0.239,MAPT were 1.000±0.071,3.297±0.243,0.497±0.029.In blank group B,overexpression group B and recovery group B,the expressions of MAPT-IT1 were 1.000±0.081,5.716±0.309,5.288±0.176,miR-181a-5p were 1.000±0.024,0.291±0.022,3.648±0.073,and MAPT were 1.000±0.054,3.309±0.177,0.883±0.075.After overexpression of MAPT-IT1,the expression of miR-181a-5p was down-regulated,while the expression of MAPT was significantly increased(P<0.001),and the proliferation and invasion ability of MDA-MB-231 and MCF-7 cells were significantly decreased(P<0.05).Re-expression of miR-181a-5p down-regulated MAPT and promoted cell pr
作者
刘洋
丁祥玺
李姿员
Liu Yang;Ding Xiangxi;Li Ziyuan(Henan Pingdingshan Maternal and Child Health Care Hospital,Pingdingshan 467000,China)
出处
《中华内分泌外科杂志》
CAS
2022年第2期164-169,共6页
Chinese Journal of Endocrine Surgery
基金
河南省科技研发专项(192102310074)。
