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NSPc1 多梳蛋白复合物在神经胶质瘤 H4 细胞中与 lncRNAs 结合并相互作用。

NSPc1 polycomb protein complex binds and cross‑talks to lncRNAs in glioma H4 cells.

机构信息

Department of Biochemistry and Molecular Biology, Logistics University of Chinese People's Armed Police Force, Tianjin 300309, P.R. China.

Jiangsu Provincial Corps Hospital of Chinese People's Armed Police Force, Tianjin 300309, P.R. China.

出版信息

Oncol Rep. 2019 Apr;41(4):2575-2584. doi: 10.3892/or.2019.7000. Epub 2019 Feb 5.

Abstract

Recently, emerging evidence shows that a number of long non‑coding RNAs (lncRNAs) recruit polycomb group (PcG) proteins to specific chromatin loci to silence relevant gene expression. In the present study, we provided evidence that lncRNA candidates, selected by bioinformatic analysis and nervous system polycomb 1 (NSPc1), a key polycomb repressive complex 1 (PRC1) member, were highly expressed in glioma H4 cells in contrast to that noted in non‑cancerous cells. RNA binding protein immunoprecipitation (RIP) assays demonstrated that metastasis associated lung adenocarcinoma transcript 1 (MALAT1), SOX2 overlapping transcript (SOX2OT) and maternally expressed 3 (MEG3) among the 8 candidates bound to the NSPc1 protein complex in glioma H4 cells. Furthermore, overexpression of NSPc1 caused a decrease in the expression of MALAT1 and MEG3 and increased expression of SOX2OT, while NSPc1 downregulation caused the levels of all three genes to increase. Meanwhile, suppression of the expression of MALAT1 increased the expression levels of mRNA and protein of NSPc1, whereas downregulation of the expression of SOX2OT decreased NSPc1 expression. Moreover, a significant decrease in cell growth and increased cell apoptosis were observed in the transfected H4 cells by MTT assay and flow cytometric analysis. The results showed that the reduced co‑expression between NSPc1 and MALAT1/SOX2OT decreased the proliferation and promoted the death of H4 cells more obviously than the respectively decrease in expression of NSPc1, MALAT1 and SOX2OT. Remarkably, the influence of a simultaneously decreased expression of NSPc1 and SOX2OT on promoting cell apoptosis was more obvious than the total effect of the separate downregulation of NSPc1 and SOX2OT on accelerating cell death. However, that impact was partially counteracted in the silencing of the co‑expression of MALAT1 and NSPc1. Furthermore, they cooperated to affect transcription of p21 and OCT4.Briefly, these data suggest NSPc1 polycomb protein complex binding and cross‑talk to lncRNAs in glioma H4 cells, offering new insight into the important function of polycomb protein complex and lncRNA interactions in glioma cells and provide a novel view of potential biomarkers and targets for the diagnosis and therapy of glioma.

摘要

最近的研究证据表明,许多长链非编码 RNA(lncRNA)募集多梳组(PcG)蛋白到特定染色质位置,以沉默相关基因的表达。在本研究中,我们提供的证据表明,通过生物信息学分析和神经系统多梳 1(NSPc1)选择的 lncRNA 候选物,NSPc1 是关键的多梳抑制复合物 1(PRC1)成员,在神经胶质瘤 H4 细胞中高表达,而在非癌性细胞中表达较低。RNA 结合蛋白免疫沉淀(RIP)分析表明,在神经胶质瘤 H4 细胞中,8 个候选物中的转移相关肺腺癌转录物 1(MALAT1)、SOX2 重叠转录物(SOX2OT)和母系表达 3(MEG3)与 NSPc1 蛋白复合物结合。此外,NSPc1 的过表达导致 MALAT1 和 MEG3 的表达减少,SOX2OT 的表达增加,而 NSPc1 的下调导致这三个基因的水平均增加。同时,抑制 MALAT1 的表达增加了 NSPc1 的 mRNA 和蛋白表达水平,而下调 SOX2OT 的表达则降低了 NSPc1 的表达。此外,通过 MTT 检测和流式细胞术分析,在转染的 H4 细胞中观察到细胞生长显著减少和细胞凋亡增加。结果表明,与 NSPc1 和 MALAT1/SOX2OT 的共表达减少相比,NSPc1 和 MALAT1/SOX2OT 的表达分别降低,H4 细胞的增殖和促进死亡更为明显。值得注意的是,同时降低 NSPc1 和 SOX2OT 的表达对促进细胞凋亡的影响比单独下调 NSPc1 和 SOX2OT 对加速细胞死亡的总效应更为明显。然而,沉默 MALAT1 和 NSPc1 的共表达部分抵消了这种影响。此外,它们合作影响 p21 和 OCT4 的转录。简而言之,这些数据表明 NSPc1 多梳蛋白复合物与神经胶质瘤 H4 细胞中的 lncRNA 结合和相互作用,为多梳蛋白复合物和 lncRNA 相互作用在神经胶质瘤细胞中的重要功能提供了新的见解,并为神经胶质瘤的诊断和治疗提供了新的潜在生物标志物和靶点。

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