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环状RNA circDNM3OS作为miR-145-5p的海绵，通过上调MORC2促进胆管癌生长和谷氨酰胺代谢。

Circular RNA circDNM3OS Functions as a miR-145-5p Sponge to Accelerate Cholangiocarcinoma Growth and Glutamine Metabolism by Upregulating MORC2.

作者信息

Su Yongfeng, Yu Ting, Wang Yaqi, Huang Xianming, Wei Xiaoyong

机构信息

Department of General Oncology, Jiangxi Provincial Cancer Hospital, Nanchang, Jiangxi, 330029, People's Republic of China.

Department of Pathology, Jiangxi Provincial Cancer Hospital, Jiangxi, 330029, People's Republic of China.

出版信息

Onco Targets Ther. 2021 Feb 17;14:1117-1129. doi: 10.2147/OTT.S289241. eCollection 2021.

DOI:10.2147/OTT.S289241
PMID:33628035
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7898209/
Abstract

BACKGROUND

Cholangiocarcinoma (CCA) is the second most common liver malignant tumor. CircRNA hsa_circ_0005230 (circDNM3OS) has been reported to exert an oncogenic role in CCA. However, the mechanisms related to circDNM3OS in CAA progression have not been fully elucidated.

METHODS

The expression of circDNM3OS, microRNA (miR)-145-5p, and MORC2 (MORC Family CW-Type Zinc Finger 2) mRNA were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, colony formation, migration, invasion, and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8), colony formation, transwell, wound-healing, and flow cytometry assays. The levels of glutamine, α-KG (α-ketoglutarate), and ATP (adenosine triphosphate) were detected using commercial kits. The relationship between circDNM3OS or MORC2 and miR-145-5p was verified by dual-luciferase reporter and/or RNA immunoprecipitation (RIP) assays. Protein level of MORC2 was measured by Western blotting. The role of circDNM3OS in CCA growth was verified by xenograft experiment.

RESULTS

CircDNM3OS and MORC2 were upregulated while miR-145-5p was downregulated in CCA tissues and cells. Inhibition of circDNM3OS reduced xenograft tumor growth in vivo and constrained proliferation, colony formation, migration, invasion, induced apoptosis, and reduced glutamine metabolism of CCA cells in vitro. CircDNM3OS sponged miR-145-5p to elevate MORC2 expression. MiR-145-5p silencing overturned circDNM3OS knockdown-mediated influence on malignancy and glutamine metabolism of CCA cells. Also, MORC2 overexpression reversed the repressive impact of miR-145-5p mimic on malignancy and glutamine metabolism of CCA cells.

CONCLUSION

CircDNM3OS facilitates CCA growth and glutamine metabolism by regulating the miR-145-5p/MORC2 pathway, offering a novel mechanism to understand the progression of CCA.

摘要

背景

胆管癌（CCA）是第二常见的肝脏恶性肿瘤。据报道，环状RNA hsa_circ_0005230（circDNM3OS）在CCA中发挥致癌作用。然而，circDNM3OS在CCA进展中的相关机制尚未完全阐明。

方法

通过定量实时聚合酶链反应（qRT-PCR）分析circDNM3OS、微小RNA（miR）-145-5p和MORC2（MORC家族CW型锌指蛋白2）mRNA的表达。通过细胞计数试剂盒-8（CCK-8）、集落形成、Transwell、伤口愈合和流式细胞术分析评估细胞增殖、集落形成、迁移、侵袭和凋亡。使用商业试剂盒检测谷氨酰胺、α-酮戊二酸（α-KG）和三磷酸腺苷（ATP）的水平。通过双荧光素酶报告基因和/或RNA免疫沉淀（RIP）分析验证circDNM3OS或MORC2与miR-145-5p之间的关系。通过蛋白质印迹法检测MORC2的蛋白水平。通过异种移植实验验证circDNM3OS在CCA生长中的作用。

结果

在CCA组织和细胞中，circDNM3OS和MORC2上调，而miR-145-5p下调。抑制circDNM3OS可降低体内异种移植肿瘤的生长，并在体外抑制CCA细胞的增殖、集落形成、迁移、侵袭，诱导凋亡并降低谷氨酰胺代谢。circDNM3OS通过海绵吸附miR-145-5p来提高MORC2的表达。miR-145-5p沉默可逆转circDNM3OS敲低介导的对CCA细胞恶性程度和谷氨酰胺代谢的影响。此外，MORC2过表达可逆转miR-145-5p模拟物对CCA细胞恶性程度和谷氨酰胺代谢的抑制作用。

结论

circDNM3OS通过调节miR-145-5p/MORC2途径促进CCA生长和谷氨酰胺代谢，为理解CCA的进展提供了一种新机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b2c70e2c375e/OTT-14-1117-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/e7ceec0989e7/OTT-14-1117-g0001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b6fa901b67b7/OTT-14-1117-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/c01e57c7f9a3/OTT-14-1117-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b2c70e2c375e/OTT-14-1117-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/e7ceec0989e7/OTT-14-1117-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b5fb816d16db/OTT-14-1117-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/d297a7a26eb9/OTT-14-1117-g0003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b6fa901b67b7/OTT-14-1117-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/c01e57c7f9a3/OTT-14-1117-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1a9/7898209/b2c70e2c375e/OTT-14-1117-g0007.jpg

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