CLDN5 affects lncRNAs acting as ceRNA dynamics contributing to regulating blood-brain barrier permeability in tumor brain metastasis
CLDN5 affects lncRNAs acting as ceRNA dynamics contributing to regulating blood-brain barrier permeability in tumor brain metastasis
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DOI：
10.3892/or.2018.6208
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发表时间：
2018-03-01
期刊：
影响因子：
4.2
通讯作者：
Jia, Wang
中科院分区：
文献类型：
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作者：
Ma, Shun-Chang;Li, Qi;Jia, Wang
血脑屏障（BBB）构成了一个有效的组织的紧密连接，限制了肿瘤的交付到大脑。BBB中的主要紧密连接蛋白是紧密连接蛋白-5（CLDN 5），但其作用机制在很大程度上仍然未知。长链非编码RNA（longnon-codingRNA，lncRNA）在多种肿瘤中异常表达，其中一些lncRNA在调节血脑屏障通透性中起重要作用，并参与肿瘤脑转移。具体地，lncRNA可以作为竞争性内源RNA（ceRNA）起作用。在此，我们研究了ceRNA失调是否与人脑血管内皮hCMEC/D3细胞中CLDN 5水平的改变相关。使用Affyscore Human Transcriptome Array 2.0和Affyscore GeneChip miRNA 4.0 Array检测pLL3.7-CLDN 5转染和pLL3.7对照hCMEC/D3细胞中2，578个miRNA、22，829个lncRNA和44，699个mRNA的表达水平。构建miRNA-lncRNA-mRNA相互作用网络。共发现41个miRNA、954个lncRNA和222个mRNA在CLDN 5过表达组和对照组之间差异表达。基于miRNA-lncRNA-mRNA相互作用网络，鉴定了148种作为ceRNA的lncRNA。通过GO和通路分析确定差异mRNA在网络中的功能。这些调控ceRNA的生物学功能主要包括紧密连接、粘着斑、细胞间粘附、细胞生长和凋亡等。通过定量实时RT-PCR实验验证鉴定的对CLDN 5过表达的hCMEC/D3细胞特异的miRNA、lncRNA和mRNA的集合。我们的研究预测了脑血管内皮细胞中与CLDN 5改变相关的大量ceRNA的生物学功能。我们的数据表明，这些失调的ceRNA与高CLDN 5水平相结合，可以作为预防脑转移形成的有用靶标。需要进一步研究以确定这些ceRNA在促进CLDN 5在脑肿瘤屏障中的功能中的作用。
The blood-brain barrier (BBB) constitutes an efficient organization of tight junctions that limits the delivery of tumor to the brain. The principal tight junction protein in BBB is claudin-5 (CLDN5), but its mechanism of action remains largely unknown. Long non-coding RNAs (lncRNAs) are aberrantly expressed in many cancers, some lncRNAs play key roles in regulating BBB permeability and are involved in tumor brain metastasis. In particular, lncRNAs can function as competing endogenous RNAs (ceRNAs). Herein, we investigated whether ceRNA dysregulation is associated with alterations of the level of CLDN5 in human brain vascular endothelial hCMEC/D3 cells. The Affymetrix Human Transcriptome Array 2.0 and Affymetrix GeneChip miRNA 4.0 Array were used to detect the expression levels of 2,578 miRNAs, 22,829 lncRNAs, and 44,699 mRNAs in pLL3.7-CLDN5-transfected and pLL3.7 control hCMEC/D3 cells. The distinctly expressed miRNAs, lncRNAs, and mRNAs were subjected to construction of miRNA-lncRNA-mRNA interaction network. A total of 41 miRNAs, 954 lncRNAs, and 222 mRNAs were found to be differentially expressed between the CLDN5-overexpressing and control group. 148 lncRNA acting as ceRNAs were identified based on the miRNA-lncRNA-mRNA interaction network. The function of differential mRNA in the network was determined by GO and pathway analysis. The potential roles of the 27 ceRNAs were revealed, the possible biology functions of these regulatory ceRNAs mainly included tight junction, focal adhesion, cell-cell adhesion, cell growth and apoptosis. The identified sets of miRNAs, lncRNAs and mRNAs specific to CLDN5-overexpressing hCMEC/D3 cells were verified by quantitative real-time RT-PCR experiment. Our study predicts the biological functions of a multitude of ceRNAs associated with the alteration of CLDN5 in brain vascular endothelial cells. Our data suggest that these dysregulated ceRNAs, in conjunction with the high CLDN5 levels, could serve as useful targets of prevention of brain metastasis formation. Further studies are warranted to determine the role of these ceRNAs in facilitating the function of CLDN5 in brain-tumor barrier.