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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。
OBJECTIVE: To screen and analyze the differential expression profiles of microRNAs (miRNAs) between laryngeal carcinoma and surrounding normal laryngeal mucosa, and to provide clues for further study on the relationship between miRNA and laryngeal carcinoma. Methods: A total of 42 pairs of laryngeal tissues and adjacent normal laryngeal mucosa specimens were collected and 10 pairs of specimens were randomly selected for miRNA microarray gene chip analysis. 32 pairs of specimens were selected for real-time quantitative PCR (qRT-PCR) to obtain laryngeal cancer MiRNA differential expression profiles in tissues. The effects of miR-125a-5p on the proliferation of laryngeal carcinoma Hep2 cells were detected by MTT assay and clonogenic assay. Results: Six miRNAs such as let-7f-5p, miR-10a-5p, miR-125a-5p, miR-144-3p, miR-195-5p and miR- PCR expression were significantly down-regulated. Compared with the control group, the proliferation of Hep2 cells transfected with miR-125a-mimics group was inhibited, while the proliferation of Hep2 cells transfected with miR-125a-inhibitor group was enhanced. Conclusion: Gene microarrays are consistent with qRT-PCR results; miRNAs are differentially expressed between laryngeal carcinoma and normal laryngeal mucosa. The differential expression of these miRNAs may be related to the pathogenesis and invasion of laryngeal carcinoma. miR-125a can inhibit the proliferation of laryngeal Hep2 cells, which may serve as a new target for biological treatment of laryngeal cancer.