肺炎支原体肺炎患儿血液中差异表达miRNAs的筛选及鉴定
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国家自然科学基金(青年科学基金项目)(81200012)


Identification of differentially expressed bloody miRNAs in children with Mycoplasma pneumoniae pneumonia
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    摘要:

    目的:筛选鉴定与肺炎支原体肺炎相关的特异性miRNAs。方法:收集肺炎支原体肺炎患儿的血液,通过miRNA芯片技术分析肺炎支原体肺炎患儿和正常人血液淋巴细胞中miRNAs的表达谱变化,通过RT-PCR方法验证芯片检测结果的准确性,确认与肺炎支原体肺炎相关的特异性miRNAs。利用靶基因软件预测各自可能的靶基因,利用RT-PCT验证mRNA表达差异。结果:肺炎支原体肺炎患儿血液淋巴细胞中筛选出表达上调的miRNA 105个,表达下调的133个。其中上调(大于5倍)的30个,上调(大于10倍)的10个,下调(小于0.5倍)的133个(P<0.05)。选取表达差异显著的miR-20a-3p、miR-100-5p、miR-93-5p以及let-7b-5进行后续RT-PCR的验证,结果与芯片一致;Bcl-2、IGF-1R、PTEN、TGFβR1 mRNA表达有不同程度的降低。结论:miRNAs在肺炎支原体肺炎患儿血液淋巴细胞中存在差异表达,而这些差异表达的miRNAs及其潜在靶基因可能参与肺炎支原体导致的感染与免疫反应。

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    Objective:To explore related miRNAs to Mycoplasma pneumoniae(Mp) caused respiratory infections and immune reaction. Methods:Blood of children with Mp pneumonia(MPP) was collected. By miRNA microarray technology,miRNAs expression differences in lymphocytes were analysed between MPP and control group. RT-PCR was used to verify the accuracy of microarray results. Specific miRNAs associated with MPP were explored. Target gene software was used to predict the possible target genes,and RT-PCT was used to verify the mRNA expression difference. Results:Compared with normal group,in MPP group 105 miRNAs increased(with 30 more than 5 fold change,10 more than 10) and 133 decreased (with 133 less than 0.5 fold) (P<0.05). Significantly differentially expressed miRNAs:miR-20a-3p,miR-100-5p,miR-93-5p,and let-7b-5p increased detected by RT-PCR,and were consistent with the array results. As potential target genes,Bcl-2,IGF-1R,PTEN,and TGFβR1 mRNA expression decreased. Conclusion:miRNAs differential expressed in blood lymphocytes in children with MPP,and the difference may be involved in infection and the immune response caused by Mp.

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张佳敏,周 瑶,莫珍珍,刘 峰,唐 珩,赵德育.肺炎支原体肺炎患儿血液中差异表达miRNAs的筛选及鉴定[J].南京医科大学学报(自然科学版),2017,(5):620-624

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  • 收稿日期:2016-05-17
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  • 在线发布日期: 2017-06-01
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