miR-122-5p通过靶向Circ_MTM1在HBV相关肝纤维化中的分子机制研究
Molecular Mechanism of miR-122-5p in HBV-Associated Liver Fibrosis by Targeting Circ_MTM1
DOI: 10.12677/HJBM.2024.141008, PDF,    科研立项经费支持
作者: 姚福纯, 蔡 艳, 黄俊文, 王振生, 王海鑫, 刘贵旻：锦州医科大学，基础医学院，基础医学系，辽宁 锦州；李 彬*：锦州医科大学，基础医学院，病原生物学教研室，辽宁 锦州
关键词: 微小核糖核酸122-5p (miR-122-5p)；Circ_MTM1；乙型肝炎病毒(HBV)；肝纤维化；miR-122-5p； Circ_MTM1； Hepatitis B Virus； Liver Fibrosis
摘要: 目的：研究微小核糖核酸microRNA-122-5p (miR-122-5p)对HBV感染相关肝纤维化的作用及分子机制。方法：分别用采用HBsAg、HBeAg酶联免疫吸附剂测定(ELISA)试剂盒和实时荧光定量聚合酶链反应(RT-qPCR)检测HBV表面抗原(HBsAg)、抗原(HBeAg)水平以及HBV DNA、HBV共价闭合环状DNA (cccDNA)水平，并采用蛋白质印迹法(Western blot)检测相应蛋白水平。RT-qPCR检测人肝星状细胞(LX-2细胞)和HBV相关肝纤维化组织细胞中Circ_MTM1、miR-122-5p的表达情况。应用生物信息学软件StarBaseV3.0预测miR-122-5p的可能靶基因，并应用双荧光素酶检测方法判定miR-122-5p对其靶基因Circ_MTM1表达的调节作用。结果：miR-122-5p在HBV相关性肝纤维化组织和细胞中低表达，而Circ_MTM1在HBV相关性肝纤维化组织和细胞中高表达。miR-122-5p靶向下调Circ_MTM1的表达，通过下调Circ_MTM1可抑制HBV相关肝纤维化的进程。结论：miR-122-5p与Circ_MTM1在HBV相关肝纤维化中存在靶向关系，Circ_MTM1通过调控肝纤维化相关蛋白的水平在HBV相关肝纤维化进程中起到启动子作用。
Abstract: Objective: To study the effect of microRNA-122-5p (miR-122-5p) on infection-related liver fibrosis and its molecular mechanism. Methods: HBsAg and HBeAg enzyme-linked immunosorbent assay (ELISA) kit and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) were used to detect HBV surface antigen (HBsAg), antigen (HBeAg) levels, HBV DNA and HBV covalent closed circular DNA (cccDNA) levels, respectively. The corresponding protein levels were detected by Western blot. The expression of Circ_MTM1 and miR-122-5p in human hepatic stellate cells (LX-2 cells) and HBV-related hepatic fibrosis tissue was detected by RT-qPCR. Bioinformatics software StarBaseV3.0 was used to predict the possible target genes of miR-122-5p, and dual luciferase assay was used to determine the regulatory effect of miR-122-5p on the expression of its target gene Circ_MTM1. Results: The expression of miR-122-5p was low in HBV-associated liver fibrosis tissues and cells, while the expression of Circ_MTM1 was high in HBV-associated liver fibrosis tissues and cells. The expression of Circ_MTM1 is down-regulated by miR-122-5p, and the process of HBV-related liver fibrosis can be inhibited by down-regulating Circ_MTM1. Conclusion: There is a targeting relationship between miR-122-5p and Circ_MTM1 in HBV-related liver fibrosis, and Circ_MTM1 plays a promoter role in the process of HBV-related liver fibrosis by regulating the level of liver fibrosis-related proteins.
文章引用：姚福纯, 李彬, 蔡艳, 黄俊文, 王振生, 王海鑫, 刘贵旻. miR-122-5p通过靶向Circ_MTM1在HBV相关肝纤维化中的分子机制研究[J]. 生物医学, 2024, 14(1): 72-80. https://doi.org/10.12677/HJBM.2024.141008

参考文献

[1] Pollicino, T. and Caminiti, G. (2021) HBV-Integration Studies in the Clinic: Role in the Natural History of Infection. Viruses, 13, Article 368. [Google Scholar] [CrossRef] [PubMed]
[2] Lin, M.H., Li, H.Q., Zhu, L., Su, H.Y., Peng, L.S., Wang, C.Y., He, C.P., Liang, X.E. and Wang, Y. (2022) Liver Fibrosis in the Natural Course of Chronic Hepatitis B Viral Infection: A Systematic Review with Meta-Analysis. Digestive Diseases and Sciences, 67, 2608-2626. [Google Scholar] [CrossRef] [PubMed]
[3] Dawood, R.M., El-Meguid, M.A., Salum, G.M. and El Awady, M.K. (2020) Key Players of Hepatic Fibrosis. Journal of Interferon & Cytokine Research, 40, 472-489. [Google Scholar] [CrossRef] [PubMed]
[4] Kisseleva, T. and Brenner, D. (2021) Molecular and Cellular Mechanisms of Liver Fibrosis and Its Regression. Nature Reviews Gastroenterology & Hepatology, 18, 151-166. [Google Scholar] [CrossRef] [PubMed]
[5] Saliminejad, K., Khorram Khorshid, H.R., Soleymani Fard, S. and Ghaffari, S.H. (2019) An Overview of microRNAs: Biology, Functions, Therapeutics, and Analysis Methods. Journal of Cellular Physiology, 234, 5451-5465. [Google Scholar] [CrossRef] [PubMed]
[6] Bartel, D.P. (2004) MicroRNAs: Genomics, Biogenesis, Mechanism, and Function. Cell, 116, 281-297. [Google Scholar] [CrossRef] 
[7] Ingenito, F., Roscigno, G., Affinito, A., et al. (2019) The Role of Exo-miRNAs in Cancer: A Focus on Therapeutic and Diagnostic Applications. International Journal of Molecular Sciences, 20, Article 4687. [Google Scholar] [CrossRef] [PubMed]
[8] Liu, Y.H., Liu, J.L., Wang, Z., Zhu, X.H., Chen, X.B. and Wang, M.Q. (2019) MiR-122-5p Suppresses Cell Proliferation, Migration and Invasion by Targeting SATB1 in Nasopharyngeal Carcinoma. European Review for Medical and Pharmacological Sciences, 23, 622-629.
[9] Li, D., Chen, J., Yun, C., Li, X. and Huang, Z. (2022) MiR-122-5p Regulates the Pathogenesis of Childhood Obesity by Targeting CPEB1. Obesity Research & Clinical Practice, 16, 206-213. [Google Scholar] [CrossRef] [PubMed]
[10] Ren, P., Wu, N.A., Fu, S., Wang, W., Li, Q.I. and Cheng, Q. (2023) miR-122-5p Restrains Pancreatic Cancer Cell Growth and Causes Apoptosis by Negatively Regulating ASCT2. Anticancer Research, 43, 4379-4388. [Google Scholar] [CrossRef] [PubMed]
[11] Zhang, L., Wang, Y., Sun, J., Ma, H. and Guo, C. (2019) LINC00205 Promotes Proliferation, Migration and Invasion of HCC Cells by Targeting miR-122-5p. Pathology—Research and Practice, 215, Article ID: 152515. [Google Scholar] [CrossRef] [PubMed]
[12] Memczak, S., Jens, M., Elefsinioti, A., Torti, F., Krueger, J., Rybak, A., et al. (2013) Circular RNAs Are a Large Class of Animal RNAs with Regulatory Potency. Nature, 495, 333-338. [Google Scholar] [CrossRef] [PubMed]
[13] Kristensen, L.S., Jakobsen, T., Hager, H. and Kjems, J. (2022) The Emerging Roles of circRNAs in Cancer and Oncology. Nature Reviews Clinical Oncology, 19, 188-206. [Google Scholar] [CrossRef] [PubMed]
[14] Liu, Y., Wang, L. and Liu, W. (2022) Roles of circRNAs in the Tumorigenesis and Metastasis of HCC: A Mini Review. Cancer Management and Research, 14, 1847-1856. [Google Scholar] [CrossRef] 
[15] 林树文. CircRPS16通过miR-876-5p上调SPINK1的表达从而促进肝细胞肝癌的增殖和侵袭[D]: [博士学位论文]. 广州: 南方医科大学, 2023.
[16] Huang, G., Liang, M., Liu, H., Huang, J., Li, P., Wang, C., Zhang, Y., Lin, Y. and Jiang, X. (2020) CircRNA Hsa_circRNA_104348 Promotes Hepatocellular Carcinoma Progression through Modulating miR-187-3p/RTKN2 Axis and Activating Wnt/β-Catenin Pathway. Cell Death & Disease, 11, Article No. 1065. [Google Scholar] [CrossRef] [PubMed]
[17] 田艳茹. miR-133b与慢性乙型肝炎肝纤维化相关性研究[J]. 陕西医学杂志, 2023, 52(4): 448-451.
[18] 谢俊锋. CircRNA97通过miRNA-146b-5p/HIPK1轴逆转肝纤维化的作用及机制研究[D]: [博士学位论文]. 南昌: 南昌大学, 2023.
[19] 林嘉宜, 娄安妮, 李旭. 脂多糖刺激巨噬细胞分泌含miR-155-5p的外泌体促进肝星状细胞的活化及迁移[J]. 南方医科大学学报, 2023, 43(6): 994-1001.
[20] 王傲. MicroRNA-195-3p通过抑制PTEN的表达促进肝星状细胞活化及肝纤维化的发展[D]: [硕士学位论文]. 合肥: 安徽医科大学, 2023.
[21] Zhang, L., Chen, Y., Zhang, L.J., Wang, M., Chang, D.L., Wan, W.W., Zhang, B.X., Zhang, W.G. and Chen, X.P. (2019) HBV Induces Different Responses of the Hepatocytes and Oval Cells during HBV-Related Hepatic Cirrhosis. Cancer Letters, 443, 47-55. [Google Scholar] [CrossRef] [PubMed]
[22] Verrier, E.R., Ligat, G., Heydmann, L., Doernbrack, K., Miller, J., Maglott-Roth, A., Jühling, F., El Saghire, H., Heuschkel, M.J., Fujiwara, N., Hsieh, S.Y., Hoshida, Y., Root, D.E., Felli, E., Pessaux, P., Mukherji, A., Mailly, L., Schuster, C., Brino, L., Nassal, M. and Baumert, T.F. (2022) Cell-Based cccDNA Reporter Assay Combined with Functional Genomics Identifies YBX1 as HBV cccDNA Host Factor and Antiviral Candidate Target. Gut, 72, 1745-1757. [Google Scholar] [CrossRef] [PubMed]
[23] Martinez, M.G., Smekalova, E., Combe, E., Gregoire, F., Zoulim, F. and Testoni, B. (2022) Gene Editing Technologies to Target HBV cccDNA. Viruses, 14, Article 2654. [Google Scholar] [CrossRef] [PubMed]
[24] Li, W., Yu, X., Chen, X., Wang, Z., Yin, M., Zhao, Z. and Zhu, C. (2021) HBV Induces Liver Fibrosis via the TGF-β1/miR-21-5p Pathway. Experimental and Therapeutic Medicine, 21, Article No. 169. [Google Scholar] [CrossRef] [PubMed]
[25] Ji, D., Chen, G.F., Wang, J.C., Ji, S.H., Wu, X.W., Lu, X.J., Chen, J.L. and Li, J.T. (2020) Hsa_circ_0070963 Inhibits Liver Fibrosis via Regulation of miR-223-3p and LEMD3. Aging, 12, 1643-1655. [Google Scholar] [CrossRef] [PubMed]
[26] 朱慧慧. 远端上游结合因子FUBP1靶向核心启动子调控HBV转录的分子机制研究[D]: [硕士学位论文]. 郑州: 河南大学, 2023.
[27] 郑晓桑, 欧启水, 刘灿. HBV核心启动子的研究进展[J]. 临床检验杂志, 2023, 41(5): 351-354.

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