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长链非编码RNA DANCR通过调控miR-185-5p/HMGA2轴促进结直肠癌进展。

Long non-coding RNA DANCR accelerates colorectal cancer progression via regulating the miR-185-5p/HMGA2 axis.

作者信息

Lu Weiqun, Huang Zhiliang, Wang Jia, Liu Haiying

机构信息

Department of Gastrointestinal Surgical Oncology, Affiliated Cancer Hospital and Institute of Guangzhou Medical University, Hengzhigang Road #78, Yuexiu District, Guangzhou 510095, PR China.

出版信息

J Biochem. 2022 Mar 31;171(4):389-398. doi: 10.1093/jb/mvab011.

Abstract

Long non-coding RNAs (lncRNAs) are crucial players in tumour progression. Herein, this work was designated to decipher the clinical significance, function and molecular mechanism of a lncRNA, differentiation antagonizing non-coding RNA (DANCR) in colorectal cancer (CRC). Quantitative real-time polymerase chain reaction was adopted to examine DANCR, miR-185-5p and HMGA2 mRNA expressions in CRC tissues and cells. Both gain-of-function and loss-of-function cell models for DANCR were established, and then MTT, wound healing and Transwell, flow cytometry assays were carried out to detect the proliferation, migration, invasion, cell cycle and apoptosis of CRC cells. Dual-luciferase reporter gene assay and RIP assay were utilized to validate the targeting relationships between DANCR and miR-185-5p. Western blot was employed for detecting high mobility group A2 (HMGA2) expressions in CRC cells. In this study, we demonstrated that the expression of DANCR was elevated in CRC tissues and cell lines, and its high expression was significantly associated with increased TNM stage and positive lymph node metastasis. DANCR overexpression promoted CRC cell proliferation, migration, invasion and cell cycle progression, but inhibited apoptosis; while knocking down DANCR caused the opposite effects. DANCR was further identified as a molecular sponge for miR-185-5p, and DANCR could indirectly increase the expression of HMGA2 via repressing miR-185-5p. In conclusion, DANCR/miR-185-5p/HMGA2 axis participated in the progression of CRC.

摘要

长链非编码RNA(lncRNA)是肿瘤进展中的关键因子。在此,本研究旨在阐明一种lncRNA——分化拮抗非编码RNA(DANCR)在结直肠癌(CRC)中的临床意义、功能及分子机制。采用定量实时聚合酶链反应检测CRC组织和细胞中DANCR、miR-185-5p和HMGA2 mRNA的表达。建立了DANCR的功能获得和功能缺失细胞模型,然后进行MTT、伤口愈合、Transwell和流式细胞术检测,以检测CRC细胞的增殖、迁移、侵袭、细胞周期和凋亡。利用双荧光素酶报告基因检测和RNA免疫沉淀检测来验证DANCR与miR-185-5p之间的靶向关系。采用蛋白质免疫印迹法检测CRC细胞中高迁移率族蛋白A2(HMGA2)的表达。在本研究中,我们发现DANCR在CRC组织和细胞系中的表达升高,其高表达与TNM分期增加和阳性淋巴结转移显著相关。DANCR过表达促进CRC细胞增殖、迁移、侵袭和细胞周期进程,但抑制凋亡;而敲低DANCR则产生相反的效果。DANCR被进一步鉴定为miR-185-5p的分子海绵,并且DANCR可通过抑制miR-185-5p间接增加HMGA2的表达。总之,DANCR/miR-185-5p/HMGA2轴参与了CRC的进展。

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