miRNA-lncRNA影响人NK细胞与树突状细胞免疫功能和乳腺癌细胞侵袭力的分子机制  

Molecular mechanism of miRNA-lncRNA interaction affecting the immune function of human NK cells and dendritic cells and the molecular mechanism of invasiveness of breast cancer cells

作　　者：李雅怡 张雨洁 卓文杏[1] LI Yayi;ZHANG Yujie;ZHUO Wenxing(Department of Medical Technique, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangdong, Guangzhou 510000,China)

机构地区：[1]广州中医药大学第一附属医院医技科,广州市510000

出　　处：《河北医药》2020年第12期1771-1774,1779,共5页Hebei Medical Journal

基　　金：广东省科技计划项目(编号:00282840133709038)。

摘　　要：目的研究miRNA-lncRNA相互作用影响人NK细胞与树突状细胞免疫功能和乳腺癌细胞侵袭力的分子机制。方法实验分为4组:乳腺癌组织,正常组织,乳腺癌细胞系,正常细胞系;通过RT-qPCR检测LncRNA(MEG3)分别在4个试验组的表达;通过CCK-8检测NK细胞与树突状细胞活力;通过Transwell测定法测量乳腺癌细胞迁移和侵袭能力;通过荧光素酶活性检测miRNA(miR-21)和MEG3之间的关系;通过蛋白质免疫检测miR-21模拟物转染或MEG3转染后,NK细胞免疫功能标记蛋白(CD56)、DC免疫功能标记蛋白(CD80)和细胞侵袭标记蛋白(MMP-2和MMP-9)的表达量。结果MEG3在乳腺癌组织和细胞中下调表达(P<0.05);MEG3的过表达抑制NK细胞与树突状细胞活力以及乳腺癌细胞迁移和侵袭能力(P<0.05);过表达的miR-21导致MEG3-WT荧光素酶活性降低,但对MEG3-mut报告基因的荧光素酶活性无明显影响;过表达的MEG3减弱了miR-21模拟物对CD56、CD80、MMP-2和MMP-9蛋白的表达(P<0.05)。结论miRNA-lncRNA相互作用在人NK细胞与树突状细胞免疫功能和乳腺癌细胞侵袭进展中的调节作用,并为乳腺癌细胞治疗提供了新的潜在治疗策略。Objective To investigate the molecular mechanism of miRNA-lncRNA interaction affecting the immune function of human NK cells and dendritic cells and the molecular mechanism of invasiveness of breast cancer cells.Methods The experiment was divided into four groups:breast cancer tissue,normal tissue,breast cancer cell line,normal cell line.The expression levels of LncRNA(MEG3)were detected by RT-qPCR in the four groups,and the vitalities of NK cells and dendritic cells were detected by CCK-8.The abilities of migration and invasion of breast cancer cells were detected by Transwell assay,and the correlation between miRNA(miR-21)and MEG3 was analyzed by means of luciferase activity detection.After miR-21 mimic transfection or MEG3 transfection detected by protein immunoassay,the expression levels of NK cell immune function marker protein(CD56),DC immunological function marker protein(CD80)and cell invasion marker proteins(MMP-2 and MMP-9)were detected.Results The expression levels of MEG3 in breast cancer tissues and cells were significantly down-regulated(P<0.05),and the overexpression of MEG3 significantly inhibited viability and the ability of migration and invasion of breast cancer cells(P<0.05).The overexpression of miR-21 decreased the activity of MEG3-WT luciferase significantly,however,which had no significant effects on the luciferase activity of the MEG3-mut reporter gene,moreover,the overexpression of MEG3 decreased the expressions of CD56,CD80,MMP-2 and MMP-9 proteins by miR-21 mimics(P<0.05).Conclusion The miRNA-lncRNA interaction can regulate the immune function of human NK cells and dendritic cells in the progression of breast cancer cell invasion,which provides an new potential therapeutic strategy for the treatment of breast cancer.

关 键 词：MIRNA lncRNA 自然杀伤细胞 树突状细胞 乳腺癌细胞 

分 类 号：R737.9[医药卫生—肿瘤]