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目的:探讨hsa-miR-143/145基因簇在人食管鳞状细胞癌(ESCC)细胞株中的表达,并对其生物学特征以及功能进行预测分析。方法:通过实时荧光定量PCR法,检测人ESCC细胞株KYSE-150和正常食管上皮细胞株Het-1A中hsa-miR-143/145基因簇的相对含量;应用miRBase获取并分析hsa-miR-143/145基因簇序列特征并比较其同源性大小;应用TargetScan 6.1、PicTar及miRanda预测hsa-miR-143/145基因簇的靶基因,取三者预测结果的交集,并结合已证实的靶基因进行功能注释(GO)和通路富集分析。结果:人ESCC细胞株KYSE-150中hsa-miR-143表达水平为0.000 2±0.000 5,显著低于正常食管上皮细胞Het-1A的0.000 6±0.000 7,t=10.301,P=0.001;hsa-miR-145的表达水平为0.000 5±0.000 1,显著低于正常食管上皮细胞Het-1A的0.001 2±0.000 2,t=6.139,P=0.004。应用3种靶标预测软件预测hsa-miR-143和hsa-miR-145的靶基因,取交集有119个基因,结合已证实的靶标基因,共224个。hsa-miR-143/145基因簇在21个物种上都显示了很好的同源性,其靶基因显著富集在癌症通路、与癌症发生相关的信号通路以及心肌通路中。结论:Hsa-miR-143/145基因簇可能参与ESCC的发病机制,其预测靶基因集合富集于多个生物学过程并且显著富集于癌症通路,为后续Hsa-miR-143/145基因簇生物学功能的研究奠定了基础。
Objective: To investigate the expression of hsa-miR-143/145 gene cluster in human esophageal squamous cell carcinoma (ESCC) cell lines and to predict its biological characteristics and function. Methods: The relative content of hsa-miR-143/145 cluster in human ESCC cell line KYSE-150 and normal esophageal epithelial cell line Het-1A was detected by real-time fluorescence quantitative PCR. MiR- / 145 gene cluster and compare their homology size. TargetScan 6.1, PicTar and miRanda were used to predict the target gene of hsa-miR-143/145 cluster, and the intersection of the three predicted results was obtained. Combined with the confirmed target gene Functional annotation (GO) and pathway enrichment analysis. Results: The expression level of hsa-miR-143 in human ESCC cell line KYSE-150 was 0.000 2 ± 0.000 5, which was significantly lower than that in normal esophageal epithelial Het-1A 0.000 6 ± 0.000 7, t = 10.301, P = 0.001; hsa The expression level of miR-145 was 0.000 5 ± 0.000 1, which was significantly lower than that of normal esophageal epithelial cells Het-1A 0.001 ± 0.000 2, t = 6.139, P = 0.004. Three target prediction softwares were used to predict the target genes of hsa-miR-143 and hsa-miR-145. A total of 119 genes were selected and 224 genes were confirmed with the target genes. The hsa-miR-143/145 gene cluster showed good homology in all 21 species, and its target genes were significantly enriched in cancer pathways, signal pathways involved in cancer development, and myocardial pathways. Conclusions: The Hsa-miR-143/145 gene cluster may be involved in the pathogenesis of ESCC. The predicted Hsa-miR-143/145 cluster enriches in multiple biological processes and is significantly enriched in the cancer pathway. The study of biological function lays the foundation.