MicroRNA‑21 regulates the expression of BTG2 in HepG2 liver cancer cells

Mao,Bijing; Xiao,He; Zhang,Zhimin; Wang,Dong; Wang,Ge

doi:10.3892/mmr.2015.4051

MicroRNA‑21 regulates the expression of BTG2 in HepG2 liver cancer cells

Authors:
- Bijing Mao
- He Xiao
- Zhimin Zhang
- Dong Wang
- Ge Wang
View Affiliations

Affiliations: Cancer Center, Institute of Surgical Research, Daping Hospital, Third Military Medical University, Chongqing 400042, P.R. China, Department of Oncology, Wuhan General Hospital of Guangzhou Command, People's Liberation Army, Wuhan, Hubei 430070, P.R. China
Published online on: July 7, 2015 https://doi.org/10.3892/mmr.2015.4051
Pages: 4917-4924
Copyright: © Mao 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

B‑cell translocation gene 2 (BTG2) is a tumor suppressor gene, which belongs to the anti‑proliferation gene family. Our previous study demonstrated that microRNA (miR)‑21 and the expression of BTG2 were negatively correlated during hepatocarcinogenesis. The aim of the present study was to investigate the effects of miR‑21 on the growth and progression of liver cancer cells, and to determine the underlying mechanism. A luciferase reporter assay was used to demonstrate that the BTG2 gene was a direct target of miR‑21. In addition, the effects of miR‑21 on cell growth and gene expression in HepG2 human hepatocellular carcinoma (HCC) cells were analyzed using reverse transcription‑quantitative polymerase chain reaction, western blotting, an MTT assay, flow cytometry, a Transwell invasion assay and a wound healing assay. The expression levels of miR‑21 in the HepG2 cells were significantly higher, compared with those in L02 normal liver cells. The expression levels of BTG2 in liver cancer cell lines (HepG2 and Huh7) were significantly lower, compared with that in the L02 cells. These results suggested that BTG2 was the direct target gene of miR‑21. The protein expression levels of BTG2 were inhibited by high expression levels of miR‑21, and increased by inhibition of the expression of miR‑21 in the HepG2 cells. Inhibition of miR‑21 reduced cell proliferation and invasion, and increased the rate of apoptosis in the HepG2 cells. These results indicated that miR‑21 regulates cell proliferation, invasion, migration and apoptosis in HepG2 cells, which may be associated with its effects on the expression of BTG2. The results of the present study may provide a basis for targeting the miR‑21/BTG2 interaction for the treatment of HCC.

View Figures

View References

1	Saito Y, Hibino S and Saito H: Alterations of epigenetics and microRNA in hepatocellular carcinoma. Hepatol Res. 44:31–42. 2014. View Article : Google Scholar
2	Liu J, Liu X, Cui F, Chen G, Guan Y and He J: The efficacy of the inhalation of an aerosolized Group A streptococcal preparation in the treatment of lung cancer. Chin J Cancer Res. 24:346–352. 2012. View Article : Google Scholar
3	Skinner HD and Komaki R: Proton radiotherapy in the treatment of lung cancer. Transl Cancer Res. 1:264–270. 2012.
4	Quintavalle C and Condorelli G: Dulanermin in cancer therapy: Still much to do. Transl Lung Cancer Res. 1:158–159. 2012.PubMed/NCBI
5	Yamashita S, Yamamoto H, Mimori K, Nishida N, Takahashi H, Haraguchi N, Tanaka F, Shibata K, Sekimoto M, Ishii H, et al: MicroRNA-372 is associated with poor prognosis in colorectal cancer. Oncology. 82:205–212. 2012.PubMed/NCBI
6	Weiland M, Gao XH, Zhou L and Mi QS: Small RNAs have a large impact: Circulating microRNAs as biomarkers for human diseases. RNA Biol. 9:850–859. 2012. View Article : Google Scholar : PubMed/NCBI
7	Tjensvoll K, Svendsen KN, Reuben JM, Oltedal S, Gilje B, Smaaland R and Nordgård O: miRNA expression profiling for identification of potential breast cancer biomarkers. Biomarkers. 17:463–470. 2012. View Article : Google Scholar : PubMed/NCBI
8	Papaconstantinou I, Karakatsanis A, Gazouli M, Polymeneas G and Voros D: The role of microRNAs in liver cancer. Eur J Gastroenterol Hepatol. 24:223–228. 2012. View Article : Google Scholar : PubMed/NCBI
9	Zhang B, Pan X, Cobb GP and Anderson TA: microRNAs as oncogenes and tumor suppressors. Dev Biol. 302:1–12. 2007. View Article : Google Scholar
10	Iorio MV and Croce CM: MicroRNAs in cancer: Small molecules with a huge impact. J Clin Oncol. 27:5848–5856. 2009. View Article : Google Scholar : PubMed/NCBI
11	Hu QY, Jiang H, Su J and Jia YQ: MicroRNAs as biomarkers for hepatocellular carcinoma: A diagnostic meta-analysis. Clin Lab. 59:1113–1120. 2013.PubMed/NCBI
12	Pass HI: Biomarkers and prognostic factors for mesothelioma. Ann Cardiothorac Surg. 1:449–456. 2012.
13	Coppola V, Musumeci M, Patrizii M, Cannistraci A, Addario A, Maugeri-Saccà M, Biffoni M, Francescangeli F, Cordenonsi M, Piccolo S, et al: BTG2 loss and miR-21 upregulation contribute to prostate cell transformation by inducing luminal markers expression and epithelial-mesenchymal transition. Oncogene. 32:1843–1853. 2013. View Article : Google Scholar
14	Niu J, Shi Y, Tan G, Yang CH, Fan M, Pfeffer LM and Wu ZH: DNA damage induces NF-κB-dependent microRNA-21 up-regulation and promotes breast cancer cell invasion. J Biol Chem. 287:21783–21795. 2012. View Article : Google Scholar : PubMed/NCBI
15	Zhang BG, Li JF, Yu BQ, Zhu ZG, Liu BY and Yan M: microRNA-21 promotes tumor proliferation and invasion in gastric cancer by targeting PTEN. Oncol Rep. 27:1019–1026. 2012.PubMed/NCBI
16	Shibuya H, Iinuma H, Shimada R, Horiuchi A and Watanabe T: Clinicopathological and prognostic value of microRNA-21 and microRNA-155 in colorectal cancer. Oncology. 79:313–320. 2010. View Article : Google Scholar
17	Zhang JG, Wang JJ, Zhao F, Liu Q, Jiang K and Yang GH: MicroRNA-21 (miR-21) represses tumor suppressor PTEN and promotes growth and invasion in non-small cell lung cancer (NSCLC). Clin Chim Acta. 411:846–852. 2010. View Article : Google Scholar : PubMed/NCBI
18	Gao J, Zhang Q, Xu J, Guo L and Li X: Clinical significance of serum miR-21 in breast cancer compared with CA153 and CEA. Chin J Cancer Res. 25:743–748. 2013.
19	Petrović N, Mandušić V, Stanojević B, Lukić S, Todorović L, Roganović J and Dimitrijević B: The difference in miR-21 expression levels between invasive and non-invasive breast cancers emphasizes its role in breast cancer invasion. Med Oncol. 31:8672014. View Article : Google Scholar
20	Li T, Leong MH, Harms B, Kennedy G and Chen L: MicroRNA-21 as a potential colon and rectal cancer biomarker. World J Gastroenterol. 19:5615–5621. 2013. View Article : Google Scholar : PubMed/NCBI
21	Liu J, Zhu H, Yang X, Ge Y, Zhang C, Qin Q, Lu J, Zhan L, Cheng H and Sun X: MicroRNA-21 is a novel promising target in cancer radiation therapy. Tumour Biol. 35:3975–3979. 2014. View Article : Google Scholar : PubMed/NCBI
22	Li L, Zhou L, Li Y, Lin S and Tomuleasa C: MicroRNA-21 stimulates gastric cancer growth and invasion by inhibiting the tumor suppressor effects of programmed cell death protein 4 and phosphatase and tensin homolog. J BUON. 19:228–236. 2014.PubMed/NCBI
23	Liu M, Wu H, Liu T, Li Y, Wang F, Wan H, Li X and Tang H: Regulation of the cell cycle gene, BTG2, by miR-21 in human laryngeal carcinoma. Cell Res. 19:828–837. 2009. View Article : Google Scholar : PubMed/NCBI
24	Lim IK: TIS21 (/BTG2/PC3) as a link between ageing and cancer: Cell cycle regulator and endogenous cell death molecule. J Cancer Res Clin Oncol. 132:417–426. 2006. View Article : Google Scholar : PubMed/NCBI
25	Melamed J, Kernizan S and Walden PD: Expression of B-cell translocation gene 2 protein in normal human tissues. Tissue Cell. 34:28–32. 2002. View Article : Google Scholar : PubMed/NCBI
26	Wei S, Hao C, Li X, Zhao H, Chen J and Zhou Q: Effects of BTG2 on proliferation inhibition and anti-invasion in human lung cancer cells. Tumour Biol. 33:1223–1230. 2012. View Article : Google Scholar : PubMed/NCBI
27	Wagener N, Bulkescher J, Macher-Goeppinger S, Karapanagiotou-Schenkel I, Hatiboglu G, Abdel-Rahim M, Abol-Enein H, Ghoneim MA, Bastian PJ, Müller SC, et al: Endogenous BTG2 expression stimulates migration of bladder cancer cells and correlates with poor clinical prognosis for bladder cancer patients. Br J Cancer. 108:973–982. 2013. View Article : Google Scholar : PubMed/NCBI
28	Choi KS, Kim JY, Lim SK, Choi YW, Kim YH, Kang SY, Park TJ and Lim IK: TIS21(/BTG2/PC3) accelerates the repair of DNA double strand breaks by enhancing Mre11 methylation and blocking damage signal transfer to the Chk2(T68)-p53(S20) pathway. DNA Repair (Amst). 11:965–975. 2012. View Article : Google Scholar
29	Zhang YJ, Wei L, Liu M, Li J, Zheng YQ, Gao Y and Li XR: BTG2 inhibits the proliferation, invasion, and apoptosis of MDA-MB-231 triple-negative breast cancer cells. Tumour Biol. 34:1605–1613. 2013. View Article : Google Scholar : PubMed/NCBI
30	Zhang Z, Chen C, Wang G, Yang Z, San J, Zheng J, Li Q, Luo X, Hu Q, Li Z and Wang D: Aberrant expression of the p53-inducible antiproliferative gene BTG2 in hepatocellular carcinoma is associated with overexpression of the cell cycle-related proteins. Cell Biochem Biophys. 61:83–91. 2011. View Article : Google Scholar : PubMed/NCBI
31	Zhang ZM, Wang G, Yang ZX, Shan JL, Chen C, Jin F, Xu W, Li Q, Luo XZ, Wang D and Li ZP: The expression of B-cell translocation gene 2 in diethylnitrosamine-induced primary hepatocellular carcinoma rat model. Zhonghua Gan Zang Bing Za Zhi. 17:107–111. 2009.In Chinese. PubMed/NCBI
32	Yang CH, Yue J, Pfeffer SR, Handorf CR and Pfeffer LM: MicroRNA miR-21 regulates the metastatic behavior of B16 melanoma cells. J Biol Chem. 286:39172–39178. 2011. View Article : Google Scholar : PubMed/NCBI
33	Takahashi F, Chiba N, Tajima K, Hayashida T, Shimada T, Takahashi M, Moriyama H, Brachtel E, Edelman EJ, Ramaswamy S and Maheswaran S: Breast tumor progression induced by loss of BTG2 expression is inhibited by targeted therapy with the ErbB/HER inhibitor lapatinib. Oncogene. 30:3084–3095. 2011. View Article : Google Scholar : PubMed/NCBI
34	Zhang L, Huang H, Wu K, Wang M and Wu B: Impact of BTG2 expression on proliferation and invasion of gastric cancer cells in vitro. Mol Biol Rep. 37:2579–2586. 2010. View Article : Google Scholar
35	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
36	Krek A, Grün D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, MacMenamin P, da Piedade I, Gunsalus KC, Stoffel M and Rajewsky N: Combinatorial microRNA target predictions. Nat Genet. 37:495–500. 2005. View Article : Google Scholar : PubMed/NCBI
37	Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP and Burge CB: Prediction of mammalian microRNA targets. Cell. 115:787–798. 2003. View Article : Google Scholar : PubMed/NCBI
38	Nair VS, Maeda LS and Ioannidis JP: Clinical outcome prediction by microRNAs in human cancer: A systematic review. J Natl Cancer Inst. 104:528–540. 2012. View Article : Google Scholar : PubMed/NCBI
39	Ross SA and Davis CD: MicroRNA, nutrition, and cancer prevention. Adv Nutr. 2:472–485. 2011. View Article : Google Scholar :
40	Li Q, Wang G, Shan JL, Yang ZX, Wang HZ, Feng J, Zhen JJ, Chen C, Zhang ZM, Xu W, et al: MicroRNA-224 is upregulated in HepG2 cells and involved in cellular migration and invasion. J Gastroenterol Hepatol. 25:164–171. 2010. View Article : Google Scholar
41	Li Q, Ding C, Chen C, Zhang Z, Xiao H, Xie F, Lei L, Chen Y, Mao B, Jiang M, et al: miR-224 promotion of cell migration and invasion by targeting Homeobox D 10 gene in human hepatocellular carcinoma. J Gastroenterol Hepatol. 29:835–842. 2014. View Article : Google Scholar
42	Zhu S, Si ML, Wu H and Mo YY: MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem. 282:14328–14336. 2007. View Article : Google Scholar : PubMed/NCBI
43	Ryu MS, Lee MS, Hong JW, Hahn TR, Moon E and Lim IK: TIS21/BTG2/PC3 is expressed through PKC-delta pathway and inhibits binding of cyclin B1-Cdc2 and its activity, independent of p53 expression. Exp Cell Res. 299:159–170. 2004. View Article : Google Scholar : PubMed/NCBI