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长链非编码RNA TUG1通过调控急性髓系白血病中的miR-193a-5p/Rab10轴来调节细胞活力和死亡。

LncRNA TUG1 Regulates Cell Viability and Death by Regulating miR-193a-5p/Rab10 Axis in Acute Myeloid Leukemia.

作者信息

Li Qun, Wang Jianmin

机构信息

Department of PICU, First People's Hospital of Shangqiu City, Shangqiu, Henan Province, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Feb 13;13:1289-1301. doi: 10.2147/OTT.S234935. eCollection 2020.

DOI:10.2147/OTT.S234935
PMID:32103996
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7025684/
Abstract

BACKGROUND

Acute myeloid leukemia (AML) is a serious threat to human health. Long non-coding RNA (lncRNA) Taurine-Upregulated Gene1 (TUG1) has been reported to participate in the development and progression of several cancers, including AML. Herein, we aimed to investigate the pathognomonic role of TUG1 in AML cells and its potential mechanistic pathway.

METHODS

Quantitative real-time PCR (qRT-PCR) assay was applied to detect the expression levels of lncRNA TUG1, miR-193a-5p and Rab10 in AML bone marrow and cell lines. The CCK-8 assay was conducted to assess the cell viability of AML HL-60 and NB4 cells and cell apoptotic assay was performed to assess the cell death. Dual-luciferase reporter assay was carried out to clarify the relationships among TUG1, miR-193a-5p and Rab10. Also, the protein level of Rab10 was examined by Western blot assay.

RESULTS

LncRNA TUG1 was up-regulated in AML bone marrow and cells. Functional analysis showed that the silencing of TUG1 suppressed cell viability, while promoted cell death in AML HL-60 and NB4 cells. TUG1 targeted miR-193a-5p and negatively regulated miR-193a-5p expression. Overexpressed miR-193a-5p resulted in the decrease of cell viability and the increase in the cell death in AML cells. Restoration experiments proved that TUG1 regulated the cell viability and death of AML cells through regulating the miR-193a-5p/Rab10 axis. Rab10 was a direct target of miR-193a-5p and was inversely regulated by miR-193a-5p. TUG1 regulated the cell viability and death of AML cells through upregulating Rab10.

CONCLUSION

Silencing of lncRNA TUG1 induces a cytotoxic effect on AML cell lines through sponging miR-193a-5p and the suppression of Rab10.

摘要

背景

急性髓系白血病（AML）对人类健康构成严重威胁。据报道，长链非编码RNA（lncRNA）牛磺酸上调基因1（TUG1）参与了包括AML在内的多种癌症的发生和发展。在此，我们旨在研究TUG1在AML细胞中的特征性作用及其潜在的机制途径。

方法

应用定量实时PCR（qRT-PCR）检测AML骨髓和细胞系中lncRNA TUG1、miR-193a-5p和Rab10的表达水平。进行CCK-8试验评估AML HL-60和NB4细胞的细胞活力，并进行细胞凋亡试验评估细胞死亡情况。进行双荧光素酶报告基因试验以阐明TUG1、miR-193a-5p和Rab10之间的关系。此外，通过蛋白质免疫印迹试验检测Rab10的蛋白水平。

结果

lncRNA TUG1在AML骨髓和细胞中上调。功能分析表明，TUG1沉默可抑制AML HL-60和NB4细胞的细胞活力，同时促进细胞死亡。TUG1靶向miR-193a-5p并负向调节miR-19三个5p的表达。过表达miR-193a-5p导致AML细胞的细胞活力降低和细胞死亡增加。回复实验证明，TUG1通过调节miR-193a-5p/Rab10轴调节AML细胞的细胞活力和死亡。Rab10是miR-193a-5p的直接靶点，并受miR-193a-5p的反向调节。TUG1通过上调Rab10调节AML细胞的细胞活力和死亡。

结论

lncRNA TUG1的沉默通过吸附miR-193a-5p和抑制Rab10对AML细胞系诱导细胞毒性作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/f4cca1b89ec5/OTT-13-1289-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/13aabc4f862b/OTT-13-1289-g0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/67b6f018a753/OTT-13-1289-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/5bab7014c610/OTT-13-1289-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/f4cca1b89ec5/OTT-13-1289-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/13aabc4f862b/OTT-13-1289-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/9dd1a15e9ac2/OTT-13-1289-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/e2a2ca475e98/OTT-13-1289-g0003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/67b6f018a753/OTT-13-1289-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/5bab7014c610/OTT-13-1289-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1530/7025684/f4cca1b89ec5/OTT-13-1289-g0007.jpg

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