Abstract
Background Circular RNAs (circRNAs) have shown key regulatory roles in human malignancies. The working mechanism of circRNAs in hepatocellular carcinoma (HCC) remains to be elucidated. Methods Cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 5-ethynyl-20-deoxyuridine (EdU) assay, and colony formation assay. Xenograft tumor model was established to analyze the role of circ_MBNL3 on tumor growth in vivo. Results Circ_MBNL3 expression was notably down-regulated in HCC tissues and cell lines. Circ_MBNL3 overexpression suppressed the proliferation, migration, and invasion and induced the apoptosis of HCC cells. miR-873-5p directly targeted the 3’ untranslated region (3’UTR) of PHF2, and PHF2 was negatively regulated by miR-873-5p in HCC cells. miR-873-5p silencing-induced anti-tumor influences were largely reversed by the interference of PHF2 in HCC cells. Circ_MBNL3 restrained xenograft tumor growth in vivo. Conclusion Circ_MBNL3 restrained the proliferation, migration, and invasion and promoted the apoptosis of HCC cells depending on the regulation of miR-873-5p/PHF2 axis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Asafo-Agyei KO, Samant H (2021) Hepatocellular Carcinoma. In: StatPearls. StatPearls Publishing, Copyright © 2021, StatPearls Publishing LLC., Treasure Island (FL)
Cannell IG, Kong YW, Bushell M (2008) How do microRNAs regulate gene expression? Biochem Soc Trans 36(Pt 6):1224–1231. https://doi.org/10.1042/bst0361224
Ding B, Lou W, Liu J, Li R, Chen J, Fan W (2019) In silico analysis excavates potential biomarkers by constructing miRNA-mRNA networks between non-cirrhotic HCC and cirrhotic HCC. Cancer Cell Int 19:186. https://doi.org/10.1186/s12935-019-0901-3
Ding B, Fan W, Lou W (2020) hsa_circ_0001955 enhances in vitro proliferation, migration, and invasion of HCC cells through miR-145-5p/NRAS Axis. Mol Therapy Nucl Acids 22:445–455. https://doi.org/10.1016/j.omtn.2020.09.007
Fabian MR, Sonenberg N, Filipowicz W (2010) Regulation of mRNA translation and stability by microRNAs. Annu Rev Biochem 79:351–379. https://doi.org/10.1146/annurev-biochem-060308-103103
Fu Y, Liu M, Li F, Qian L, Zhang P, Lv F, Cheng W, Hou R (2019) MiR-221 promotes hepatocellular carcinoma cells migration via targeting PHF2. Biomed Res Int. https://doi.org/10.1155/2019/4371405
Huang G, Liang M, Liu H, Huang J, Li P, Wang C, Zhang Y, Lin Y, Jiang X (2020a) CircRNA hsa_circRNA_104348 promotes hepatocellular carcinoma progression through modulating miR-187-3p/RTKN2 axis and activating Wnt/β-catenin pathway. Cell Death Dis 11(12):1065. https://doi.org/10.1038/s41419-020-03276-1
Huang XY, Huang ZL, Huang J, Xu B, Huang XY, Xu YH, Zhou J, Tang ZY (2020b) Exosomal circRNA-100338 promotes hepatocellular carcinoma metastasis via enhancing invasiveness and angiogenesis. J Exp Clin Cancer Res: CR 39(1):20. https://doi.org/10.1186/s13046-020-1529-9
Lee KH, Ju UI, Song JY, Chun YS (2014) The histone demethylase PHF2 promotes fat cell differentiation as an epigenetic activator of both C/EBPα and C/EBPδ. Mol Cells 37(10):734–741. https://doi.org/10.14348/molcells.2014.0180
Legnini I, Di Timoteo G, Rossi F, Morlando M, Briganti F, Sthandier O, Fatica A, Santini T, Andronache A, Wade M, Laneve P, Rajewsky N, Bozzoni I (2017) Circ-ZNF609 Is a circular RNA that can be translated and functions in myogenesis. Mol Cell 66(1):22-37.e29. https://doi.org/10.1016/j.molcel.2017.02.017
Li X, Yang L, Chen LL (2018) The biogenesis, functions, and challenges of circular RNAs. Mol Cell 71(3):428–442. https://doi.org/10.1016/j.molcel.2018.06.034
Li G, Xu Y, Wang S, Yan W, Zhao Q, Guo J (2019) MiR-873-5p inhibits cell migration, invasion and epithelial-mesenchymal transition in colorectal cancer via targeting ZEB1. Pathol Res Pract 215(1):34–39. https://doi.org/10.1016/j.prp.2018.10.008
Li D, Zhang J, Li J (2020) Role of miRNA sponges in hepatocellular carcinoma. Clinica Chimica Acta Int J Clin Chem 500:10–19. https://doi.org/10.1016/j.cca.2019.09.013
Li J, Bao S, Wang L, Wang R (2021) CircZKSCAN1 suppresses hepatocellular carcinoma tumorigenesis by regulating miR-873-5p/downregulation of deleted in liver cancer 1. Dig Dis Sci. https://doi.org/10.1007/s10620-020-06789-z
Liu Z, Yu Y, Huang Z, Kong Y, Hu X, Xiao W, Quan J, Fan X (2019) CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression. Cell Death Dis 10(12):900. https://doi.org/10.1038/s41419-019-2089-9
Lou W, Liu J, Ding B, Chen D, Xu L, Ding J, Jiang D, Zhou L, Zheng S, Fan W (2019) Identification of potential miRNA-mRNA regulatory network contributing to pathogenesis of HBV-related HCC. J Transl Med 17(1):7. https://doi.org/10.1186/s12967-018-1761-7
Luo J, Zhu H, Jiang H, Cui Y, Wang M, Ni X, Ma C (2018) The effects of aberrant expression of LncRNA DGCR5/miR-873-5p/TUSC3 in lung cancer cell progression. Cancer Med 7(7):3331–3341. https://doi.org/10.1002/cam4.1566
Panda AC (2018) Circular RNAs Act as miRNA sponges. Adv Exp Med Biol 1087:67–79. https://doi.org/10.1007/978-981-13-1426-1_6
Qiu L, Xu H, Ji M, Shang D, Lu Z, Wu Y, Tu Z, Liu H (2019) Circular RNAs in hepatocellular carcinoma: Biomarkers, functions and mechanisms. Life Sci 231:116660. https://doi.org/10.1016/j.lfs.2019.116660
Ren Z, Liu X, Si Y, Yang D (2020) Long non-coding RNA DDX11-AS1 facilitates gastric cancer progression by regulating miR-873-5p/SPC18 axis. Artificial Cells, Nanomed Biotechnol 48(1):572–583. https://doi.org/10.1080/21691401.2020.1726937
Salamo O, Mortaz E, Mirsaeidi M (2018) Noncoding RNAs: new players in pulmonary medicine and sarcoidosis. Am J Respir Cell Mol Biol 58(2):147–156. https://doi.org/10.1165/rcmb.2017-0196TR
Su Y, Lv X, Yin W, Zhou L, Hu Y, Zhou A, Qi F (2019) CircRNA Cdr1as functions as a competitive endogenous RNA to promote hepatocellular carcinoma progression. Aging 11(19):8183–8203. https://doi.org/10.18632/aging.102312
Szabo L, Salzman J (2016) Detecting circular RNAs: bioinformatic and experimental challenges. Nat Rev Genet 17(11):679–692. https://doi.org/10.1038/nrg.2016.114
Wang YG, Wang T, Ding M, Xiang SH, Shi M, Zhai B (2019a) hsa_circ_0091570 acts as a ceRNA to suppress hepatocellular cancer progression by sponging hsa-miR-1307. Cancer Lett 460:128–138. https://doi.org/10.1016/j.canlet.2019.06.007
Wang L, Long H, Zheng Q, Bo X, Xiao X, Li B (2019b) Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression. Mol Cancer 18(1):119. https://doi.org/10.1186/s12943-019-1046-7
Wang Z, Liu W, Wang C, Ai Z (2020) miR-873-5p inhibits cell migration and invasion of papillary thyroid cancer via regulation of CXCL16. Onco Targets Ther 13:1037–1046. https://doi.org/10.2147/ott.s213168
Xiong DD, Dang YW, Lin P, Wen DY, He RQ, Luo DZ, Feng ZB, Chen G (2018) A circRNA-miRNA-mRNA network identification for exploring underlying pathogenesis and therapy strategy of hepatocellular carcinoma. J Transl Med 16(1):220. https://doi.org/10.1186/s12967-018-1593-5
Yang Y, Gao X, Zhang M, Yan S, Sun C, Xiao F, Huang N, Yang X, Zhao K, Zhou H, Huang S, Xie B, Zhang N (2018) Novel Role of FBXW7 Circular RNA in repressing glioma tumorigenesis. J Natl Cancer Inst 110(3):304–315. https://doi.org/10.1093/jnci/djx166
Yang JD, Hainaut P, Gores GJ, Amadou A, Plymoth A, Roberts LR (2019) A global view of hepatocellular carcinoma: trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol 16(10):589–604. https://doi.org/10.1038/s41575-019-0186-y
Yu J, Xu QG, Wang ZG, Yang Y, Zhang L, Ma JZ, Sun SH, Yang F, Zhou WP (2018) Circular RNA cSMARCA5 inhibits growth and metastasis in hepatocellular carcinoma. J Hepatol 68(6):1214–1227. https://doi.org/10.1016/j.jhep.2018.01.012
Yu J, Yang M, Zhou B, Luo J, Zhang Z, Zhang W, Yan Z (2019) CircRNA-104718 acts as competing endogenous RNA and promotes hepatocellular carcinoma progression through microRNA-218–5p/TXNDC5 signaling pathway. Clinical Sci 133(13):1487–1503. https://doi.org/10.1042/cs20190394
Zhang Y, Zhang C, Zhao Q, Wei W, Dong Z, Shao L, Li J, Wu W, Zhang H, Huang H, Liu F, Jin S (2019) The miR-873/NDFIP1 axis promotes hepatocellular carcinoma growth and metastasis through the AKT/mTOR-mediated Warburg effect. Am J Cancer Res 9(5):927–944
Acknowledgements
Not applicable.
Funding
None.
Author information
Authors and Affiliations
Contributions
JC designed and performed the research; ML and RW analyzed the data; LP wrote the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
10528_2022_10295_MOESM1_ESM.tif
Supplementary file1 Circ_MBNL3 knockdown elevates the malignant potential of HCC cells. (A-H) Hep3B and Huh1 cell lines stably transfected with sh-NC or sh-circ_MBNL3 were used to perform RT-qPCR and a series of functional experiments. (A and B) RT-qPCR was conducted to detect the expression of circ_MBNL3 and MBNL3 mRNA in HCC cells. Unpaired Student’s t-test was used to analyze the differences. (C and D) MTT assay was performed to analyze the proliferation ability of HCC cells. Unpaired Student’s t-test was used to analyze the differences. (E) EdU assay was carried out to analyze the proliferation ability of HCC cells. Unpaired Student’s t-test was used to analyze the differences. (F) Colony formation assay was performed to analyze the proliferation ability of HCC cells. Unpaired Student’s t-test was used to analyze the differences. (G and H) Cell migration and invasion abilities were analyzed by transwell assays. Unpaired Student’s t-test was used to analyze the differences. *P<0.05. (TIF 853 kb)
10528_2022_10295_MOESM2_ESM.tif
Supplementary file2 Circ_MBNL3 knockdown-mediated pro-tumor effects are overturned by silencing miR-873-5p. (A-G) HCC cells were divided into the following four groups: sh-NC, sh-circ_MBNL3, sh-circ_MBNL3 + anti-miR-NC, and sh-circ_MBNL3 + anti-miR-873-5p. (A) RT-qPCR was conducted to detect the expression of miR-873-5p in HCC cells. One-way ANOVA followed by a Tukey’s test was used to analyze the differences. (B-E) Cell proliferation ability was analyzed by MTT, EdU, and colony formation assays. One-way ANOVA followed by a Tukey’s test was used to analyze the differences. (F and G) Cell migration and invasion capacities were assessed by transwell assays. One-way ANOVA followed by a Tukey’s test was used to analyze the differences. *P<0.05. (TIF 1028 kb)
10528_2022_10295_MOESM3_ESM.tif
Supplementary file3 Circ_MBNL3 knockdown promotes xenograft tumor growth in vivo. (A) Hep3B cell line stably expressing sh-NC or sh-circ_MBNL3 was established, and the knockdown efficiency of circ_MBNL3 was assessed by RT-qPCR. Unpaired Student’s t-test was used to analyze the differences. (B) Tumor volume was monitored every 5 d via length×width2×0.5. Unpaired Student’s t-test was used to analyze the differences. (C) After inoculation for 27 d, all nude mice were killed and xenograft tumors were weighed. Unpaired Student’s t-test was used to analyze the differences. (D-F) RT-qPCR and western blot assay were conducted to measure the expression of circ_MBNL3, miR-873-5p, and PHF2 mRNA and protein. *P<0.05. (TIF 432 kb)
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Peng, L., Chen, J., Li, M. et al. Circ_MBNL3 Restrains Hepatocellular Carcinoma Progression by Sponging miR-873-5p to Release PHF2. Biochem Genet 61, 1015–1034 (2023). https://doi.org/10.1007/s10528-022-10295-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10528-022-10295-4