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LncRNA NUTM2A-AS1 aggravates the progression of hepatocellular carcinoma by activating the miR-186-5p/KLF7-mediated Wnt/beta-catenin pathway

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Abstract

Emerging evidence has uncovered that noncoding RNAs (ncRNAs) contribute to the development of hepatocellular carcinoma (HCC). Nevertheless, the functions of the majority of long ncRNAs (lncRNAs) in HCC are unknown. Here, we intend to probe the function of lncRNA NUTM2A-AS1 in the evolvement of HCC and the related mechanism. Expression levels of lncRNA NUTM2A-AS1, miR-186-5p and KLF7 mRNA in HCC tissues and adjacent non-tumor tissues were monitored. Gain- or loss-of-function assays were utilized to investigate the biological functions of lncRNA NUTM2A-AS1, miR-186-5p and KLF7 in HCC cell lines (including HCCLM3 and Huh7). Western blot was implemented for the detection of the epithelial–mesenchymal transition (EMT)-related proteins (including E-cadherin, Vimentin and Snail), KLF7, Wnt, β-catenin, and stemness-related proteins (Nanog, OCT4, YKL40, and CD133). Furthermore, the targeted associations between lncRNA NUTM2A-AS1, miR-186-5p, and KLF7 were verified by bioinformatics analysis, dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. As a result, lncRNA NUTM2A-AS1 and KLF7 profiles were heightened in the HCC tissues versus adjacent normal tissues, while miR-186-5p had the opposite expression tendency. Up-regulation of lncRNA NUTM2A-AS1 was related to tumor size, advanced tumor stage, and lymph node metastasis of HCC patients. Functionally, overexpression of lncRNA NUTM2A-AS1 heightened HCC cells’ growth, invasion, EMT, and stemness and repressed their apoptosis by activating the Wnt/β-catenin pathway. In contrast, up-regulation of miR-186-5p or inhibition of KLF7 had reverse effects. In vivo, lncRNA NUTM2A-AS1 overexpression facilitated tumor growth and EMT, accompanied by declined miR-186-5p levels and enhanced KLF7 expression. The mechanistic studies revealed that miR-186-5p served as a common target of lncRNA NUTM2A-AS1 and KLF7. As hinted by the rescue experiments, NUTM2A-AS1 partly abated miR-186-5p-mediated anti-tumor effects in HCC cells, whereas KLF7 knockdown reversed the promotive effects of NUTM2A-AS1. LncRNA NUTM2A-AS1 accelerated the evolution of HCC by up-regulating the KLF7/Wnt/beta-catenin pathway through sponging miR-186-5p.

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Data availability statement

The data sets used and analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

The study was supported by The Natural Science Foundation of Hunan Province (2022JJ30545), Key special funds project of Hunan Science and Technology Department (2020SKC2009), The Scientific Research Project of Hunan Provincial Health and Family Planning Commission (20201954, A2017012, A2017015) and The Special Funding for the Construction of Innovative Provinces in Hunan (2021SK4031).

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Hengyang Medical School, The Affiliated Nanhua Hospital, University of South China, No. 336, Dongfeng South Road, Zhuhui District, Hengyang City, 421000, China

    Jianwu Long, Longfei Liu, Xiaojun Zhou & Xianzhou Lu

  2. Department of General Surgery, The First Affiliated Hospital of Soochow University, No. 899 Pinghai Road, Gusu District, Suzhou City, 215000, Jiangsu, China

    Jianwu Long & Lei Qin

  3. Hunan Provincial Clinical Research Center for Metabolic Associated Fatty Liver Disease, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang, China

    Xuefeng Yang

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  1. Jianwu Long
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  4. Xiaojun Zhou
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Contributions

Conceived and designed the experiments: LQ. Performed the experiments: JL; FL; YY. Statistical analysis: FL; XZ; XL. Wrote the paper: JL; all the authors read and approved the final manuscript.

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Correspondence to Xianzhou Lu or Lei Qin.

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The authors declare that they have no competing interests.

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This study has been approved by The Affiliated Nanhua Hospital of University of South China. All animal procedures were performed according to the instructions of the ethics committee of The Affiliated Nanhua Hospital of University of South China.

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Long, J., Liu, L., Yang, X. et al. LncRNA NUTM2A-AS1 aggravates the progression of hepatocellular carcinoma by activating the miR-186-5p/KLF7-mediated Wnt/beta-catenin pathway. Human Cell 36, 312–328 (2023). https://doi.org/10.1007/s13577-022-00802-5

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