【摘要】 目的:探讨微小RNA-448(miR-448)对口腔鳞癌细胞增殖调控作用,筛选并验证相关靶基因。方法:RT-q PCR检测miR-448在人口腔鳞癌临床标本中表达;miR-448抑制物转染口腔鳞癌细胞系Cal-27细胞,MTT实验研究miR-448抑制物对细胞增殖能力的影响;细胞划痕实验检测miR-448抑制物对细胞迁移能力的影响。3种基因预测软件筛选出miR-448的可能下游靶基因,实验验证miR-448对靶基因的调控作用。结果:RT-q PCR分析发现miR-448在15对OSCC组织内表达显著升高。与对照组相比,转染miR-448抑制物后,Cal-27细胞增殖及迁移能力明显下降;Western blot、RT-q PCR结果显示SPARCL1基因的表达量升高;荧光素酶报告基因实验确定miR-448与SPARCL1基因在3′UTR区存在位点结合。结论:miR-448在人口腔鳞癌临床标本中呈高表达。miR-448与SPARCL1-3′UTR的结合,下调SPARCL1的表达,在口腔鳞癌细胞中发挥癌基因的作用,促进肿瘤细胞的增殖与迁移。更多还原

【Abstract】 Objective:To investigate the effect of miR-448 on the proliferation of oral squamous cell carcinoma,and to screen and verify the relvelant genes. Methods:Using RT-q PCR to study the expression of miR-448 in human OSCC. Then miR-448 inhibitor was transfected into OSCC cell line Cal-27 cells,and the effects of miR-448 inhibitor on proliferation and migration of OSCC were detected by MTT assay and scratch test. In addition,we screened the target gene of miR-448 by three gene prediction software,and verify the regulation of miR-448 on target genes. Results:We found that miR-448 was significantly increased in 15 pairs of OSCC tissues by RTq PCR analysis. Compared with the control group,the proliferation and migration of Cal-27 cells were significantly decreased after transfected with miR-448 inhibitor. The expression of SPARCL1 gene was increased detected by RT-q PCR and Western blot,and the presence of site-specific binding of miR-448 to the SPARCL1 gene in the 3′ UTR region was confirmed by luciferase reporter.Conclusions:Mi R-448 is highly expressed in clinical samples of oral squamous cell carcinoma. miR-448 down-regulates the expression of SPARCL1 by binding to SPARCL1-3′ UTR,playing an oncogene role in oral squamous cell carcinoma and promoting tumor cell proliferation and migration.更多还原