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长链非编码 RNA AFAP1-AS1 通过表观遗传抑制 p21 表达预测非小细胞肺癌患者的不良预后并调控非小细胞肺癌细胞增殖。

Long noncoding RNA AFAP1-AS1 predicts a poor prognosis and regulates non-small cell lung cancer cell proliferation by epigenetically repressing p21 expression.

机构信息

Cancer Research and Biotherapy Center, the Second Affiliated Hospital of Southeast University, Nanjing, Jiangsu, People's Republic of China.

Department of Oncology, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.

出版信息

Mol Cancer. 2018 May 24;17(1):92. doi: 10.1186/s12943-018-0836-7.

DOI:10.1186/s12943-018-0836-7
PMID:29793547
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5968553/
Abstract

BACKGROUND

Mounting evidence indicates that long noncoding RNAs (lncRNAs) could play a pivotal role in cancer biology. However, the role and molecular mechanism and global genes that were mediated by lncRNA AFAP1-AS1 in non-small cell lung cancer (NSCLC) remain largely unknown.

METHODS

Expression of AFAP1-AS1 was analyzed in 92 NSCLC tissues and cell lines by Quantitative real time polymerase chain reaction (qRT-PCR). The effect of AFAP1-AS1 on proliferation was evaluated by function assays both in in vitro and in vivo. RNA-seq assays were performed after knockdown AFAP1-AS1. RNA immunoprecipitation (RIP) was performed to confirm the interaction between AFAP1-AS1 and EZH2. Chromatin immunoprecipitation (ChIP) was used to study the promoter region of p21.

RESULTS

AFAP1-AS1 expression was increased in NSCLC tissues and was correlated with clinical outcomes of NSCLC. Further experiments revealed that inhibition of its expression in NSCLC cells resulted in diminished cell growth in vitro and in vivo. RNA-seq revealed that knockdown of AFAP1-AS1 could induce the expression of p21. Mechanistic investigations found that AFAP1-AS1 could interact with EZH2 and recruit EZH2 to the promoter regions of p21, thus epigenetically repressing p21 expression.

CONCLUSIONS

Together, these results suggest that lncRNA AFAP1-AS1 may serve as a candidate prognostic biomarker and target for new therapies in human NSCLC.

摘要

背景

越来越多的证据表明，长非编码 RNA（lncRNA）可能在癌症生物学中发挥关键作用。然而，lncRNA AFAP1-AS1 在非小细胞肺癌（NSCLC）中的作用、分子机制以及介导的全局基因仍然知之甚少。

方法

通过定量实时聚合酶链反应（qRT-PCR）分析 92 例 NSCLC 组织和细胞系中 AFAP1-AS1 的表达。通过体外和体内功能测定评估 AFAP1-AS1 对增殖的影响。敲低 AFAP1-AS1 后进行 RNA 测序分析。进行 RNA 免疫沉淀（RIP）实验以证实 AFAP1-AS1 与 EZH2 之间的相互作用。采用染色质免疫沉淀（ChIP）实验研究 p21 的启动子区域。

结果

AFAP1-AS1 在 NSCLC 组织中表达增加，与 NSCLC 的临床结局相关。进一步的实验表明，抑制 NSCLC 细胞中其表达可导致体外和体内细胞生长减少。RNA 测序显示，敲低 AFAP1-AS1 可诱导 p21 的表达。机制研究发现，AFAP1-AS1 可与 EZH2 相互作用，并募集 EZH2 到 p21 的启动子区域，从而表观遗传抑制 p21 的表达。

结论

综上所述，这些结果表明 lncRNA AFAP1-AS1 可能作为人类 NSCLC 的候选预后生物标志物和新治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/678cc84042e9/12943_2018_836_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/342a3303cac6/12943_2018_836_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/648d6922d6f4/12943_2018_836_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/e7778ecc7a6e/12943_2018_836_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/f7a69b8fd21a/12943_2018_836_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/f3abd856a3d6/12943_2018_836_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/678cc84042e9/12943_2018_836_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/342a3303cac6/12943_2018_836_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/648d6922d6f4/12943_2018_836_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/e7778ecc7a6e/12943_2018_836_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/f7a69b8fd21a/12943_2018_836_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/f3abd856a3d6/12943_2018_836_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf44/5968553/678cc84042e9/12943_2018_836_Fig6_HTML.jpg

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