摘要
目的:探讨miR-124-3p在鼻咽癌中的表达及其对鼻咽癌细胞的影响及其机制。方法:收集鼻咽癌组织标本90例和慢性鼻咽炎症组织标本85例,运用qRT-PCR方法检测组织标本和CNE1、CNE2、SUNE1、HONE1、5-8F、6-10B及C666-1鼻咽癌细胞株和永生化鼻咽上皮细胞NP69中miR-124-3p的表达,脂质体介导的转染方法下调高表达miRNA-124-3p的鼻咽癌细胞株CNE2,同时上调低表达miRNA-124-3p的鼻咽癌细胞株C666-1。CCK8法、流式细胞技术、细胞划痕实验、Transwell实验和Boyden小室检测细胞增殖、凋亡、迁移及侵袭的变化;生物信息学靶基因预测miRNA-124-3的靶基因并运用荧光素酶报告实验验证,Western Blotting检测细胞中STAT3及其下游的p-STAT3、CCND2和MMP2蛋白表达水平。结果:鼻咽癌组织中miRNA-124-3p表达水平与慢性鼻咽炎症组织相比明显下调(P<0.001);miRNA-124-3p的表达水平与肿瘤大小及范围、区域淋巴结受累情况及临床分期显著相关(均P<0.001);7种鼻咽癌细胞与NP69细胞相比较,miRNA-124-3p的表达水平均显著低于NP69(均P<0.05)。miRNA-124-3p过表达后C666-1细胞增殖、凋亡、迁移和侵袭均显著低于空白对照组和mimics NC组(均P<0.05);miRNA-124-3p抑制后CNE2细胞的增殖、凋亡、迁移和侵袭均显著高于inhibitor NC组和空白对照组(均P<0.05)。转染miRNA-124-3p后C666-1细胞STAT3、p-STAT3、CCND2和MMP2的表达显著降低,抑制CNE2细胞的miRNA-124-3p表达后STAT3、p-STAT3、CCND2和MMP2的表达显著降低。结论:miR-124-3p可通过下调靶基因STAT3的表达,进而影响其下游的p-STAT3、CCND2和MMP2信号通路,从而促进鼻咽癌细胞凋亡,抑制鼻咽癌细胞的增殖、迁移及侵袭。
Objective： To investigate the effects of microRNA- 124- 3p（ miR- 124- 3p） expressions on nasopharyngeal carcinoma（ NPC） cells and its relevant mechanism. Methods： A total of 90 NPC tissues and 85 postnasal catarrh tissues were collected. Quantitative real- time polymerase chain reaction（ qRT- PCR） was used to detect tissue samples and expressions of miR- 124- 3p in CNE1,CNE2,SUNE1,HONE1,5- 8F,6- 10 B and C666- 1 NPC cell line and immortalized nasopharyngeal epithelial cells line（ NP69）. Over- expressed miRNA- 124- 3p in CNE2 was down- regulated,and low- expressed miRNA- 124- 3p in C666- 1 was up- regulated by liposome- mediated transfection. Cell Counting Kit（ CCK- 8）,flow cytometry,the scratch test,transwell migration assay and Boyden chamber assays were used to detect cell proliferation,apoptosis,migration and invasion. The target gene of miRNA-124- 3 calculated by Bioinformatics was further determined using dual luciferase system. Protein levels of signal transducers and activators of transcription 3（ STAT3）,phospho- STAT3（ p- STAT3）,mouse anti human cyclin D2（ CCND2） and matrix metalloproteinase- 2（ MMP2） were tested by Western Blotting. Results： MiRNA- 124- 3p expressions in NPC were markedly down- regulated than postnasal catarrh tissues（ P〈0. 001）; miRNA- 124-3p expressions showed closely linkage with clinical stages,regional lymph node involvement and T stages（ all P〈0. 001）. miRNA- 124- 3p expressions were lower in 7 NPC cells than NP69 cells（ all P〈0. 05）. After up- regulation of miR- 124- 3p,proliferation,apoptosis,migration and invasion ability of C666- 1 cell were suppressed; while after down- regulation of miR- 124- 3p,CNE2 cell abilities were increased（ all P〈0. 05）. STAT3,p- STAT3,CCND2 and MMP2 expressions in C666- 1 cells were decreased after transfected with miRNA- 124- 3p,and the above protein expressions in CNE2 cells were increased after inhibition of miRNA- 124- 3p（ all P〈0. 05）. Conclusion： miR- 124- 3p may negativel
出处
《现代肿瘤医学》
CAS
2016年第18期2866-2874,共9页
Journal of Modern Oncology
基金
青年科学基金项目(编号:81102057)
