Objective:To explore the molecular mechanism of microRNA -24( miR -24 ) on laryngeal squamous carcinoma( LSCC ) Hep2 cells invasion ability. Methods: Software miRanda and RNA22 were used to predict the miR - 24 targeted gene and its binding sites, and then quantitative real - time PCR and Western blot were used to detect the expression of S100A8 mRNA and protein respectively after transfection of Hep2 cells with pre - miR -24 according to the prediction result. The invasive ability of Hep2 cells were assayed by Transwell after anti - S100A8 blocked. Results: S100A8 3UTR contained a miR -24 binding site, which had a homology up to 95% with rat. In transfected cells, the increased accumulation of miR - 24 resulted in a significantly decreased S100A8 protein level ( P < 0. 05 ), but had no effect on the level of S100A8 mRNA( P > 0. 05 ). The invasion ability of Hep2 cells were statistically inhibited while compared anti - S100A8 blocked group with non - blocking group, P < 0. 05. Conclusions: miR -24 could negatively regulate the expression of S100A8 by binding to the 3UTR, which lead LSCC Hep2 cells invasion ability inhibited.%目的:探讨microRNA-24(miRNA-24)对喉鳞状细胞癌(laryngeal squamous carcinoma,LSCC)Hep2细胞侵袭影响的分子机制.方法:应用miRanda和RNA22靶基因预测软件预测miR-24的靶基因及其结合位点,根据预测结果通过转染miR-24前体上调其在Hep2细胞中的表达,应用荧光定量PCR和Western blot分别检测miR-24过表达对S100钙结合蛋白A8(S100 calcium binding protein A8,S100A8) mRNA和蛋白表达变化的影响.S100A8抗体阻断方法,检测转染miR-24前体后Hep2细胞侵袭能力的变化.结果:S100A8 3′非翻译区(3′-untranslation region,3′UTR)含有一个miR-24结合位点,而且该位点与大鼠的同源性高达95%;miR-24基因转染的Hep2细胞中miR-24 的表达显著升高,S100A8蛋白的表达显著降低,(P均<0.05),而S100A8 mRNA表达变化未见显著性差异,(P>0.05).与未阻断组相比,miR-24前体转染能明显降低阻断S100A8组的Hep2细胞侵袭能力,(P<0.05).结论:LSCC Hep2细胞中,miR-24可结合到S100A8基因的3′UTR,在转录后水平上负性调控S100A8基因的表达,从而抑制Hep2细胞侵袭.
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