喉癌组织中的microRNA差异表达谱及miR-125a-5p抑制喉癌细胞增殖的初步研究

Differential microRNA expression profile in laryngeal cancer and effect of miR-125a-5p on proliferation of laryngeal cancer cell line

目的:筛选并分析喉癌组织与周围正常喉黏膜的微小RNA(microRNAs,miRNAs)之间的表达谱差异,为进一步研究miRNA与喉癌发生、发展的关系提供线索。方法:收集喉癌组织和癌旁正常喉黏膜标本共42对,随机选取10对标本进行miRNA微阵列基因芯片分析,另选取32对标本进行实时荧光定量PCR(qRT-PCR)验证,获得喉癌组织中的miRNA差异表达谱。应用MTT法和克隆形成实验检测miR-125a-5p对喉癌Hep2细胞增殖的影响。结果:喉癌组织中的let-7f-5p、miR-10a-5p、miR-125a-5p、miR-144-3p、miR-195-5p、miR-203等6个miRNA在基因芯片以及qRT-PCR中表达均显著下调。与对照组相比,转染miR-125a-mimics组的喉癌Hep2细胞增殖能力受到抑制,而转染miR-125a-inhibitor组Hep2细胞增殖能力增强。结论:基因芯片与qRT-PCR结果一致;喉癌与正常喉黏膜之间存在明显的miRNA差异表达,这些miRNA的差异性表达可能与喉癌的发病、侵袭等相关。miR-125a可以抑制喉癌Hep2细胞的增殖,可能作为喉癌生物治疗的新靶点。

AIM： To investigate the differential microRNA expression profiles between laryngeal cancer and adjacent normal laryngeal mucosa. METHODS： Forty two pairs of laryngeal cancer tissue and adjacent normal laryngeal mucosa tissue were collected. Ten pairs of samples were used for determining microRNA expression by the method of miR- NA microarray chip. Data analysis was performed to find out the significant differential microRNA expression profile in la- ryngeal cancer, and the difference was verified by quantitative real-time PCR （qRT- PCR） analysis on another 32 pairs of samples. Methyl thiazolyl tetrazolium （MTT） assay and colony-forming assay were used to analyze the proliferation of Hep2 cells induced by miR-125a-Sp. RESULTS： Both miRNA microarray and qRT-PCR showed that the expression of let-7b 5p, miR-10a-5p, miR-125a-5p, miR-144-3p, miR-195-5p and miR-203 was down-regulated in laryngeal cancer tissues. miR-125a-Sp suppressed the proliferation of Hep2 cells. CONCLUSION： The results of microarray are accordant with those of qRT-PCR. Significant difference of miRNA expression profiles between laryngeal cancer and adjacent normal laryn- geal mucosa indicates that miRNAs may play a role in carcinogenesis and progression of laryngeal cancer, miR-125a-5p in- hibits the proliferation of Hep2 cell, indicating a novel therapeutic target against laryngeal cancer.