The topological key lncRNA H2k2 from the ceRNA network promotes mesangial cell proliferation in diabetic nephropathy via the miR-449a/b/Trim11/Mek signaling pathway
The topological key lncRNA H2k2 from the ceRNA network promotes mesangial cell proliferation in diabetic nephropathy via the miR-449a/b/Trim11/Mek signaling pathway
复制标题
ceRNA网络中的拓扑关键lncRNA H2k2通过miR-449a/b/Trim11/Mek信号通路促进糖尿病肾病肾小球系膜细胞增殖
DOI：
10.1096/fj.201900522r
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发表时间：
2019-10-01
期刊：
影响因子：
4.8
通讯作者：
Zhang, Zheng
中科院分区：
文献类型：
--
作者：
Chen, Wenyun;Peng, Rui;Zhang, Zheng
有证据表明，竞争性内源性RNA（ceRNA）网络中的长链非编码RNA（lncRNA）参与各种疾病。然而，目前对糖尿病肾病（diabetic nephropathy，DN）中ceRNA网络的研究尚不多见。在这项研究中，我们研究了在DN相关的ceRNA网络中lncRNA对系膜细胞（MC）增殖的影响。用RNA-seq检测DN与正常小鼠肾组织中lncRNA和mRNA表达的差异，用R3.4.3构建DN相关lncRNA/mRNA/microRNA（miRNA）ceRNA网络。进行计算分析，并通过生物信息学方法、实时定量PCR（qPCR）和荧光素酶测定来检测拓扑RNA之间的表达和相互作用。用5-乙炔基-2 '-脱氧尿苷（EdU）法检测高糖或低糖条件下培养的MCs的增殖能力。此外，通过EdU、流式细胞术分析和Western blot检测拓扑关键lncRNA组织相容性2 K区位点2（H2 k2）H2 k2通过miRNA（miR）-449 a/B/三联体基序11（Trim 11）/Mek信号通路对MC增殖的影响。总共有153个lncRNA，428个mRNA和2242个相互作用被包括在构建的DN相关的ceRNA网络中。在度和介数的前5%中有15个RNA。DN组MCs中lncRNA H2 k2和mRNA Trim 11表达增加，与RNA-seq和real-time qPCR的结果一致。选择在高糖培养的MC中下调的miR-449 a和miR-449 b用于进一步分析。实时定量PCR和荧光素酶检测结果显示，H2 k2-miR-449 a/b-Trim 11在MCs中存在相互作用。此外，数据显示H2 k2通过miR-449 ab/Trim 11/Mek信号通路调节MC增殖。总之，这些结果为DN相关ceRNA网络中拓扑关键lncRNA H2 k2与DN MC增殖期间miR-449 a/B/Trim 11/Mek信号通路之间的关联提供了新的见解。陈伟，彭河，巴西-地Sun，Y.，刘洪，张，L.，彭，H.，Zhang，Z.来自ceRNA网络的拓扑关键lncRNA H2 k2通过miR-449 a/B/Trim 11/Mek信号通路促进糖尿病肾病中的系膜细胞增殖
Evidence has shown that long noncoding RNAs (lncRNAs) in the competing endogenous RNA (ceRNA) network are involved in various diseases. However, there is a lack of studies of the ceRNA network in diabetic nephropathy (DN). In this study, we investigated the effect of lncRNAs on mesangial cell (MC) proliferation in DN-related ceRNA networks. Differences in lncRNA and mRNA expression between DN and normal mouse kidney tissues were detected with RNA-seq, and DN-related lncRNA/mRNA/microRNA (miRNA) ceRNA networks were constructed by R3.4.3. Computational analysis was performed, and expression and interactions between the topological RNAs were detected by bioinformatics methods, real-time quantitative PCR (qPCR), and luciferase assay. Cell proliferation ability was measured by 5-ethynyl-2'-deoxyuridine (EdU) in MCs cultured under high- or low-glucose conditions. Moreover, the effect of the topological key lncRNA histocompatibility 2 K region locus 2 (H2k2) H2k2 on MC proliferation via the miRNA (miR)-449a/b/triplet motif 11 (Trim11)/Mek signaling pathway was examined by EdU, flow cytometry analysis, and Western blot. In total, 153 lncRNAs, 428 mRNAs, and 2242 interactions were included in the constructed DN-related ceRNA network. There were 15 RNAs in the top 5% of degree and betweenness. The expression of lncRNA H2k2 and mRNA Trim11 in MCs was increased in DN, which is consistent with the results of RNA-seq and real-time qPCR in vivo and in vitro. miR-449a and miR-449b, which were down-regulated in MCs cultured with high glucose, were selected for further analysis. The results of real-time qPCR and luciferase assay revealed the lncRNA H2k2-miR-449a/b-Trim11 interaction in MCs. In addition, the data showed that H2k2 regulates MC proliferation via the miR-449ab/Trim11/Mek signaling pathway. Taken together, these results provide new insight into the association between the topological key lncRNA H2k2 in the DN-related ceRNA network and the miR-449a/b/Trim11/Mek signaling pathway during MC proliferation in DN.-Chen, W., Peng, R., Sun, Y., Liu, H., Zhang, L., Peng, H., Zhang, Z. The topological key lncRNA H2k2 from the ceRNA network promotes mesangial cell proliferation in diabetic nephropathy via the miR-449a/b/Trim11/Mek signaling pathway.