Overexpression of MiR-450 Affects the Biological Behavior of HepG2 Cells by Targeting DNMT3a

Overview

Journal Onco Targets Ther

Publisher Dove Medical Press

Specialty Oncology

Date 2019 Jul 16

PMID 31303764

Citations 4

Authors

Yan Wang

Lina Wang

Xue Yu

Jianping Duan

Affiliations

Soon will be listed here.

Abstract

Purpose: This study was designed to explore the regulation mechanism of miR-450 in the development of hepatocarcinoma, and the effects of overexpression of miR-450 on biological behaviors such as proliferation, migration, and invasion of hepatoma cells.

Methods: HepG2 cells were divided into miR-450 mimics group, miR-450 inhibitor group, miR-450 mimics NC group, miR-450 inhibitor NC group, and blank group. MTT assay was served to measure cell proliferation, and Transwell assay was used to test cell migration and invasion. Additionally, cell cycle was detected by flow cytometry and apoptosis was examined with AnnexinV-PI double staining. After the target gene of miR-450 was predicted by bioinformatics software, Western blot and dual luciferase reporter gene experiment were applied to verify the relationship between miR-450 and target gene.

Results: The MTT and Transwell assay indicated that overexpression of miR-450 inhibited the proliferation, invasion, and migration of HepG2 cells. The flow cytometry analysis showed that overexpression of miR-450 arrested the cell cycle in the G1 phase. Meanwhile, Annexin V-PI double staining assay revealed that overexpression of miR-450 promoted apoptosis of HepG2 cells. However, silencing miR-450 in HepG2 cells promoted proliferation and invasion, and reduced apoptosis. Moreover, we found that DNMT3a was the target gene of miR-450.

Conclusions: miR-450 could inhibit proliferation, invasion, and migration via regulating DNMT3a in hepatocarcinoma cells, which provided a theoretical basis for the treatment of liver cancer.

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References

Saito Y, Kanai Y, Sakamoto M, Saito H, Ishii H, Hirohashi S . Expression of mRNA for DNA methyltransferases and methyl-CpG-binding proteins and DNA methylation status on CpG islands and pericentromeric satellite regions during human hepatocarcinogenesis. Hepatology. 2001; 33(3):561-8. DOI: 10.1053/jhep.2001.22507. View

Oh B, Kim H, Park H, Shim Y, Choi J, Park C . DNA methyltransferase expression and DNA methylation in human hepatocellular carcinoma and their clinicopathological correlation. Int J Mol Med. 2007; 20(1):65-73. View

Shikauchi Y, Saiura A, Kubo T, Niwa Y, Yamamoto J, Murase Y . SALL3 interacts with DNMT3A and shows the ability to inhibit CpG island methylation in hepatocellular carcinoma. Mol Cell Biol. 2009; 29(7):1944-58. PMC: 2655625. DOI: 10.1128/MCB.00840-08. View

Zhao Z, Wu Q, Cheng J, Qiu X, Zhang J, Fan H . Depletion of DNMT3A suppressed cell proliferation and restored PTEN in hepatocellular carcinoma cell. J Biomed Biotechnol. 2010; 2010:737535. PMC: 2868982. DOI: 10.1155/2010/737535. View

Weng Z, Wang D, Zhao W, Song M, You F, Yang L . microRNA-450a targets DNA methyltransferase 3a in hepatocellular carcinoma. Exp Ther Med. 2012; 2(5):951-955. PMC: 3440703. DOI: 10.3892/etm.2011.288. View

Luo M, Weng Y, Tang J, Hu M, Liu Q, Jiang F . MicroRNA-450a-3p represses cell proliferation and regulates embryo development by regulating Bub1 expression in mouse. PLoS One. 2012; 7(10):e47914. PMC: 3478270. DOI: 10.1371/journal.pone.0047914. View

Zhao Z, Li R, Sha S, Wang Q, Mao W, Liu T . Targeting HER3 with miR-450b-3p suppresses breast cancer cells proliferation. Cancer Biol Ther. 2014; 15(10):1404-12. PMC: 4130733. DOI: 10.4161/cbt.29923. View

Jin Y, Jiang Z, Guan X, Chen Y, Tang Q, Wang G . miR-450b-5p Suppresses Stemness and the Development of Chemoresistance by Targeting SOX2 in Colorectal Cancer. DNA Cell Biol. 2016; 35(5):249-56. DOI: 10.1089/dna.2015.3120. View

Chamani F, Sadeghizadeh M, Masoumi M, Babashah S . Evaluation of MiR-34 Family and DNA Methyltransferases 1, 3A, 3B Gene Expression Levels in Hepatocellular Carcinoma Following Treatment with Dendrosomal Nanocurcumin. Asian Pac J Cancer Prev. 2016; 17(S3):219-24. DOI: 10.7314/apjcp.2016.17.s3.219. View

10.

Liu F, Yu X, Huang H, Chen X, Wang J, Zhang X . Upregulation of microRNA-450 inhibits the progression of lung cancer in vitro and in vivo by targeting interferon regulatory factor 2. Int J Mol Med. 2016; 38(1):283-90. DOI: 10.3892/ijmm.2016.2612. View

11.

Wu Y, Bu F, Yu H, Li W, Huang C, Meng X . Methylation of Septin9 mediated by DNMT3a enhances hepatic stellate cells activation and liver fibrogenesis. Toxicol Appl Pharmacol. 2016; 315:35-49. DOI: 10.1016/j.taap.2016.12.002. View

12.

Affo S, Yu L, Schwabe R . The Role of Cancer-Associated Fibroblasts and Fibrosis in Liver Cancer. Annu Rev Pathol. 2016; 12:153-186. PMC: 5720358. DOI: 10.1146/annurev-pathol-052016-100322. View

13.

Qin X, Chen J, Wu L, Liu Z . MiR-30b-5p acts as a tumor suppressor, repressing cell proliferation and cell cycle in human hepatocellular carcinoma. Biomed Pharmacother. 2017; 89:742-750. DOI: 10.1016/j.biopha.2017.02.062. View

14.

Lopane C, Agosti P, Gigante I, Sabba C, Mazzocca A . Implications of the lysophosphatidic acid signaling axis in liver cancer. Biochim Biophys Acta Rev Cancer. 2017; 1868(1):277-282. DOI: 10.1016/j.bbcan.2017.06.002. View

15.

Xie D, Yuan P, Wang D, Jin H, Chen H . Expression and prognostic significance of and in liver cancer. Oncol Lett. 2017; 14(2):2305-2309. PMC: 5530117. DOI: 10.3892/ol.2017.6423. View

16.

Ringelhan M, McKeating J, Protzer U . Viral hepatitis and liver cancer. Philos Trans R Soc Lond B Biol Sci. 2017; 372(1732). PMC: 5597741. DOI: 10.1098/rstb.2016.0274. View

17.

Castelli G, Pelosi E, Testa U . Liver Cancer: Molecular Characterization, Clonal Evolution and Cancer Stem Cells. Cancers (Basel). 2017; 9(9). PMC: 5615342. DOI: 10.3390/cancers9090127. View

18.

Akinyemiju T, Abera S, Ahmed M, Alam N, Alemayohu M, Allen C . The Burden of Primary Liver Cancer and Underlying Etiologies From 1990 to 2015 at the Global, Regional, and National Level: Results From the Global Burden of Disease Study 2015. JAMA Oncol. 2017; 3(12):1683-1691. PMC: 5824275. DOI: 10.1001/jamaoncol.2017.3055. View

19.

Zhang Z, Chen J, Xia S, Zhao H, Yang J, Zhang H . Long intergenic non-protein coding RNA 319 aggravates lung adenocarcinoma carcinogenesis by modulating miR-450b-5p/EZH2. Gene. 2018; 650:60-67. DOI: 10.1016/j.gene.2018.01.096. View

20.

Wang Y, Xie Y, Li X, Lin J, Zhang S, Li Z . MiR-876-5p acts as an inhibitor in hepatocellular carcinoma progression by targeting DNMT3A. Pathol Res Pract. 2018; 214(7):1024-1030. DOI: 10.1016/j.prp.2018.04.012. View