Association between microRNA‑527 and glypican‑3 in hepatocellular carcinoma

Nomura,Kei; Kitanaka,Akira; Iwama,Hisakazu; Tani,Joji; Nomura,Takako; Nakahara,Mai; Ohura,Kyoko; Tadokoro,Tomoko; Fujita,Koji; Mimura,Shima; Yoneyama,Hirohito; Kobara,Hideki; Morishita,Asahiro; Okano,Keiichi; Suzuki,Yasuyuki; Tsutsi,Kunihiko; Himoto,Takashi; Masaki,Tsutomu

doi:10.3892/ol.2021.12490

Association between microRNA‑527 and glypican‑3 in hepatocellular carcinoma

Authors:
- Kei Nomura
- Akira Kitanaka
- Hisakazu Iwama
- Joji Tani
- Takako Nomura
- Mai Nakahara
- Kyoko Ohura
- Tomoko Tadokoro
- Koji Fujita
- Shima Mimura
- Hirohito Yoneyama
- Hideki Kobara
- Asahiro Morishita
- Keiichi Okano
- Yasuyuki Suzuki
- Kunihiko Tsutsi
- Takashi Himoto
- Tsutomu Masaki
View Affiliations

Affiliations: Department of Gastroenterology and Neurology, Faculty of Medicine, Kagawa University, Miki‑cho, Kagawa 761‑0793, Japan, Department of Laboratory Medicine, Kawasaki Medical School, Kurashiki, Okayama 701‑0192, Japan, Information Technology Center, Kagawa University, Miki‑cho, Kagawa 761‑0793, Japan, Department of Gastroenterological Surgery, Faculty of Medicine, Kagawa University, Miki‑cho, Kagawa 761‑0793, Japan, Department of Healthy Science, Faculty of Medicine, Kagawa University, Miki‑cho, Kagawa 761‑0793, Japan, Department of Clinical Examination, Faculty of Health Sciences, Kagawa Prefectural University of Health Sciences, Takamatsu, Kagawa 761‑0123, Japan
Published online on: January 26, 2021 https://doi.org/10.3892/ol.2021.12490
Article Number: 229
Copyright: © Nomura et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to identify the specific microRNAs (miRNAs/miRs) and their corresponding target genes involved in hepatocellular carcinomas (HCCs). Microarray analysis was performed to examine the miRNA expression profiles of four paired HCC and corresponding non‑cancerous (N) liver tissues using 985 miRNA probes. The Human miRNA Target database was used to identify the target genes of differentially expressed miRNAs between the HCC and N tissues. The protein expression levels of target genes in the HCC tissues and cell lines were evaluated using western blotting. miRNA‑mediated suppression of target gene expression was evaluated by transiently transfecting the miRNA into the HCC cell lines. Of the 985 miRNAs evaluated, four miRNAs were differentially expressed (three upregulated and one downregulated miRNAs). Of these four miRNAs, miRNA‑527 was highly downregulated in the HCC tissues. Glypican‑3 (GPC‑3) was predicted as a target gene of miRNA‑527. Western blotting revealed that GPC‑3 protein is highly expressed in the HCC tissues and HCC cell lines compared with N and normal cell lines. Transfection with miR‑527 resulted in suppression of GPC‑3 protein expression in the Cos7 cells. Furthermore, transfection with miR‑527 also inhibited the intrinsic expression of GPC‑3 in the Huh‑7 cell line. This indicated that miR‑527 in the HCC tissues may be an important novel miRNA that targets the GPC‑3 gene expression. GPC‑3, whose expression is regulated by miR‑527, may be involved in the development and progression of HCC.

View Figures

View References

1	Yang JD, Hainaut P, Gores GJ, Amadou A, Plymoth A and Roberts LR: A global view of hepatocellular carcinoma: Trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol. 16:589–604. 2019. View Article : Google Scholar : PubMed/NCBI
2	Wang CH, Wey KC, Mo LR, Chang KK, Lin RC and Kuo JJ: Current trends and recent advances in diagnosis, therapy, and prevention of hepatocellular carcinoma. Asian Pac J Cancer Prev. 16:3595–3604. 2015. View Article : Google Scholar : PubMed/NCBI
3	Sherman M: Hepatocellular carcinoma: Epidemiology, surveillance, and diagnosis. Semin Liver Dis. 30:3–16. 2010. View Article : Google Scholar : PubMed/NCBI
4	Iwama H, Masaki T and Kuriyama S: Abundance of microRNA target motifs in the 3′-UTRs of 20527 human genes. FEBS Lett. 581:1805–1810. 2007. View Article : Google Scholar : PubMed/NCBI
5	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
6	Krek A, Grün D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, MacMenamin P, da Piedade I, Gunsalus KC, Stoffel M, et al: Combinatorial microRNA target predictions. Nat Genet. 37:495–500. 2005. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: Metazoan MicroRNAs. Cell. 173:20–51. 2018. View Article : Google Scholar : PubMed/NCBI
8	Tomasello L, Cluts L and Croce CM: Experimental validation of microRNA targets: Mutagenesis of binding regions. Methods Mol Biol. 1970:331–339. 2019. View Article : Google Scholar : PubMed/NCBI
9	Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST and Patel T: MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology. 133:647–658. 2007. View Article : Google Scholar : PubMed/NCBI
10	Takamizawa J, Konishi H, Yanagisawa K, Tomida S, Osada H, Endoh H, Harano T, Yatabe Y, Nagino M, Nimura Y, et al: Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64:3753–3756. 2004. View Article : Google Scholar : PubMed/NCBI
11	Varnholt H, Drebber U, Schulze F, Wedemeyer I, Schirmacher P, Dienes HP and Odenthal M: MicroRNA gene expression profile of hepatitis C virus-associated hepatocellular carcinoma. Hepatology. 47:1223–1232. 2008. View Article : Google Scholar : PubMed/NCBI
12	Visone R and Croce CM: miRNAs and cancer. Am J Pathol. 174:1131–1138. 2009. View Article : Google Scholar : PubMed/NCBI
13	Morishita A and Masaki T: miRNA in hepatocellular carcinoma. Hepatol Res. 45:128–141. 2015. View Article : Google Scholar : PubMed/NCBI
14	Calin GA, Liu CG, Sevignani C, Ferracin M, Felli N, Dumitru CD, Shimizu M, Cimmino A, Zupo S, Dono M, et al: MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc Natl Acad Sci USA. 101:11755–11760. 2004. View Article : Google Scholar : PubMed/NCBI
15	Sevignani C, Calin GA, Siracusa LD and Croce CM: Mammalian microRNAs: A small world for fine-tuning gene expression. Mamm Genome. 17:189–202. 2006. View Article : Google Scholar : PubMed/NCBI
16	Metzler M, Wilda M, Busch K, Viehmann S and Borkhardt A: High expression of precursor microRNA-155/BIC RNA in children with Burkitt lymphoma. Genes Chromosomes Cancer. 39:167–169. 2004. View Article : Google Scholar : PubMed/NCBI
17	Michael MZ, O'Connor SM, van Holst Pellekaan NG, Young GP and James RJ: Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol Cancer Res. 1:882–891. 2003.PubMed/NCBI
18	Tokarz P and Blasiak J: The role of microRNA in metastatic colorectal cancer and its significance in cancer prognosis and treatment. Acta Biochim Pol. 59:467–474. 2012. View Article : Google Scholar : PubMed/NCBI
19	Del Vescovo V and Denti MA: microRNA and lung cancer. Adv Exp Med Biol. 889:153–177. 2015. View Article : Google Scholar : PubMed/NCBI
20	Yang X, Liang L, Zhang XF, Jia HL, Qin Y, Zhu XC, Gao XM, Qiao P, Zheng Y, Sheng YY, et al: MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin-6-Stat3 pathway. Hepatology. 58:158–170. 2013. View Article : Google Scholar : PubMed/NCBI
21	Zhu Y, Lu Y, Zhang Q, Liu JJ, Li TJ, Yang JR, Zeng C and Zhuang SM: MicroRNA-26a/b and their host genes cooperate to inhibit the G1/S transition by activating the pRb protein. Nucleic Acids Res. 40:4615–4625. 2012. View Article : Google Scholar : PubMed/NCBI
22	Wang Y, Lu Y, Toh ST, Sung WK, Tan P, Chow P, Chung AYF, Jooi LLP and Lee CG: Lethal-7 is down-regulated by the hepatitis B virus × protein and targets signal transducer and activator of transcription 3. J Hepatol. 53:57–66. 2010. View Article : Google Scholar : PubMed/NCBI
23	Wang Z, Lin S, Li JJ, Xu Z, Yao H, Zhu X, Xie D, Shen Z, Sze J, Li K, et al: MYC protein inhibits transcription of the microRNA cluster MC-let-7a-1~let-7d via noncanonical E-box. J Biol Chem. 286:39703–39714. 2011. View Article : Google Scholar : PubMed/NCBI
24	Tsang WP and Kwok TT: Let-7a microRNA suppresses therapeutics-induced cancer cell death by targeting caspase-3. Apoptosis. 13:1215–1222. 2008. View Article : Google Scholar : PubMed/NCBI
25	Di Fazio P, Montalbano R, Neureiter D, Alinger B, Schmidt A, Merkel AL, Quint K and Ocker M: Downregulation of HMGA2 by the pan-deacetylase inhibitor panobinostat is dependent on hsa-let-7b expression in liver cancer cell lines. Exp Cell Res. 318:1832–1843. 2012. View Article : Google Scholar : PubMed/NCBI
26	Zhu XM, Wu LJ, Xu J, Yang R and Wu FS: Let-7c microRNA expression and clinical significance in hepatocellular carcinoma. J Int Med Res. 39:2323–2329. 2011. View Article : Google Scholar : PubMed/NCBI
27	Lan FF, Wang H, Chen YC, Chan CY, Ng SS, Li K, Xie D, He ML, Lin MC and Kung HF: Hsa-let-7g inhibits proliferation of hepatocellular carcinoma cells by downregulation of c-Myc and upregulation of p16(INK4A). Int J Cancer. 128:319–331. 2011. View Article : Google Scholar : PubMed/NCBI
28	Traister A, Shi W and Filmus J: Mammalian Notum induces the release of glypicans and other GPI-anchored proteins from the cell surface. Biochem J. 410:503–511. 2008. View Article : Google Scholar : PubMed/NCBI
29	Li N, Gao W, Zhang YF and Ho M: Glypicans as Cancer Therapeutic Targets. Trends Cancer. 4:741–754. 2018. View Article : Google Scholar : PubMed/NCBI
30	Capurro MI, Xu P, Shi W, Li F, Jia A and Filmus J: Glypican-3 inhibits Hedgehog signaling during development by competing with patched for Hedgehog binding. Dev Cell. 14:700–711. 2008. View Article : Google Scholar : PubMed/NCBI
31	Shirakawa H, Suzuki H, Shimomura M, Kojima M, Gotohda N, Takahashi S, Nakagohri T, Konishi M, Kobayashi N, Kinoshita T, et al: Glypican-3 expression is correlated with poor prognosis in hepatocellular carcinoma. Cancer Sci. 100:1403–1407. 2009. View Article : Google Scholar : PubMed/NCBI
32	Zhang J, Zhang M, Ma H, Song X, He L, Ye X and Li X: Overexpression of glypican-3 is a predictor of poor prognosis in hepatocellular carcinoma: An updated meta-analysis. Medicine (Baltimore). 97:e111302018. View Article : Google Scholar : PubMed/NCBI
33	Fujihara S, Kato K, Morishita A, Iwama H, Nishioka T, Chiyo T, Nishiyama N, Miyoshi H, Kobayashi M, Kobara H, et al: Antidiabetic drug metformin inhibits esophageal adenocarcinoma cell proliferation in vitro and in vivo. Int J Oncol. 46:2172–2180. 2015. View Article : Google Scholar : PubMed/NCBI
34	Fujita K, Iwama H, Sakamoto T, Okura R, Kobayashi K, Takano J, Katsura A, Tatsuta M, Maeda E, Mimura S, et al: Galectin-9 suppresses the growth of hepatocellular carcinoma via apoptosis in vitro and in vivo. Int J Oncol. 46:2419–2430. 2015. View Article : Google Scholar : PubMed/NCBI
35	Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
36	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
37	Towbin H, Staehelin T and Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA. 76:4350–4354. 1979. View Article : Google Scholar : PubMed/NCBI
38	Benjamini Y and Hochberg Y: Controlling the false discovery rate: A practical and powerful approach to multiple testing. J R Stat Soc B. 57:289–300. 1995.
39	Chen Y-H, Song Y, Yu YL, Cheng W and Tong X: miRNA-10a promotes cancer cell proliferation in oral squamous cell carcinoma by upregulating GLUT1 and promoting glucose metabolism. Oncol Lett. 17:5441–5446. 2019.PubMed/NCBI
40	Pal R and Greene S: microRNA-10b is overexpressed and critical for cell survival and proliferation in medulloblastoma. PLoS One. 10:e01378452015. View Article : Google Scholar : PubMed/NCBI
41	Werk AN, Bruckmueller H, Haenisch S and Cascorbi I: Genetic variants may play an important role in mRNA-miRNA interaction: Evidence for haplotype-dependent downregulation of ABCC2 (MRP2) by miRNA-379. Pharmacogenet Genomics. 24:283–291. 2014. View Article : Google Scholar : PubMed/NCBI
42	Nishida T and Kataoka H: Glypican 3-targeted therapy in hepatocellular carcinoma. Cancers (Basel). 11:E13392019. View Article : Google Scholar : PubMed/NCBI
43	Vongchan P and Linhardt RJ: Characterization of a new monoclonal anti-glypican-3 antibody specific to the hepatocellular carcinoma cell line, HepG2. World J Hepatol. 9:368–384. 2017. View Article : Google Scholar : PubMed/NCBI
44	Singh AK, Kumar R and Pandey AK: Hepatocellular Carcinoma: Causes, Mechanism of Progression and Biomarkers. Curr Chem Genomics Transl Med. 12:9–26. 2018. View Article : Google Scholar : PubMed/NCBI
45	Zhou F, Shang W, Yu X and Tian J: Glypican-3: A promising biomarker for hepatocellular carcinoma diagnosis and treatment. Med Res Rev. 38:741–767. 2018. View Article : Google Scholar : PubMed/NCBI
46	Wang L, Yao M, Pan LH, Qian Q and Yao DF: Glypican-3 is a biomarker and a therapeutic target of hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int. 14:361–366. 2015. View Article : Google Scholar : PubMed/NCBI
47	Lai JP, Sandhu DS, Yu C, Han T, Moser CD, Jackson KK, Guerrero RB, Aderca I, Isomoto H, Garrity-Park MM, et al: Sulfatase 2 up-regulates glypican 3, promotes fibroblast growth factor signaling, and decreases survival in hepatocellular carcinoma. Hepatology. 47:1211–1222. 2008. View Article : Google Scholar : PubMed/NCBI
48	Huo W, Zhu XM, Pan XY, Du M, Sun Z and Li ZM: MicroRNA-527 inhibits TGF-β/SMAD induced epithelial-mesenchymal transition via downregulating SULF2 expression in non-small-cell lung cancer. Math Biosci Eng. 16:4607–4621. 2019. View Article : Google Scholar : PubMed/NCBI