Emodin inhibits TGF-β2 by activating the FOXD3/miR‑199a axis in ovarian cancer cells in vitro

Song,Kejuan; Lv,Teng; Chen,Yulong; Diao,Yuchao; Yao,Qin; Wang,Yankui

doi:10.3892/or.2018.6301

Emodin inhibits TGF-β2 by activating the FOXD3/miR‑199a axis in ovarian cancer cells in vitro

Authors:
- Kejuan Song
- Teng Lv
- Yulong Chen
- Yuchao Diao
- Qin Yao
- Yankui Wang
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China
Published online on: March 6, 2018 https://doi.org/10.3892/or.2018.6301
Pages: 2063-2070
Copyright: © Song 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

Ovarian cancer is a highly metastatic malignancy and a leading cause of cancer-related death in postmenopausal women. Emodin is a natural anthraquinone isolated from several traditional Chinese medicines including Rhubarb and Polygonum cuspidatum. Recently, emodin was demonstrated to reduce the growth of human ovarian carcinoma cells. However, the mechanism remains unclear. In the present study, we identified that transforming growth factor (TGF)-β2 was significantly affected by emodin treatment in A2780 cells using microarray analysis. MicroRNA (miR)-199a was predicted as a potential miRNA targeting TGF-β2 by in silico prediction using TargetScan. The mRNA and protein levels of TGF-β2 were both significantly reduced by miR-199a. Spearman's correlation analysis revealed a significant correlation between the expression level of miR-199a and TGF-β2 in human ovarian cancer specimens. Silencing of miR-199a with miR-199a inhibitor significantly restored the reduction in TGF-β2 expression induced by emodin. Additionally, cell viability and colony formation of A2780 cells were markedly inhibited by emodin treatment, which was mediated by miR-199a. We analyzed the primary mature miR-199a-1 and miR-199a-2 transcripts in A2780 cells treated with emodin or dimethyl sulfoxide (DMSO) and found that only pri-miR-199a-1 was regulated by emodin. A conserved binding site of Forkhead box D3 (FOXD3) was identified within pri-miR‑199a-1. We further revealed that miR-199a expression was significantly regulated by FOXD3. Taken together, the present study demonstrated that emodin may directly promote FOXD3 expression and sequentially activates miR-199a, which in turn suppresses the expression of TGF-β2 to reduce cell viability and colony formation of A2780 cells.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
3	Dong X, Fu J, Yin X, Cao S, Li X, Lin L and Ni J: Huyiligeqi: Emodin: A review of its pharmacology, toxicity and pharmacokinetics. Phytother Res. 30:1207–1218. 2016. View Article : Google Scholar : PubMed/NCBI
4	Lin SZ, Xu JB, Ji X, Chen H, Xu HT, Hu P, Chen L, Guo JQ, Chen MY, Lu D, et al: Emodin inhibits angiogenesis in pancreatic cancer by regulating the transforming growth factor-β/drosophila mothers against decapentaplegic pathway and angiogenesis-associated microRNAs. Mol Med Rep. 12:5865–5871. 2015. View Article : Google Scholar : PubMed/NCBI
5	Guo J, Xiao B, Liu Q, Gong Z and Le Y: Suppression of C-myc expression associates with anti-proliferation of aloe-emodin on gastric cancer cells. Cancer Invest. 26:369–374. 2008. View Article : Google Scholar : PubMed/NCBI
6	Suboj P, Babykutty S, Srinivas P and Gopala S: Aloe emodin induces G2/M cell cycle arrest and apoptosis via activation of caspase-6 in human colon cancer cells. Pharmacology. 89:91–98. 2012. View Article : Google Scholar : PubMed/NCBI
7	Xie MJ, Ma YH, Miao L, Wang Y, Wang HZ, Xing YY, Xi T and Lu YY: Emodin-provoked oxidative stress induces apoptosis in human colon cancer HCT116 cells through a p53-mitochondrial apoptotic pathway. Asian Pac J Cancer Prev. 15:5201–5205. 2014. View Article : Google Scholar : PubMed/NCBI
8	Gao R, Chen R, Cao Y, Wang Y, Song K, Zhang Y and Yang J: Emodin suppresses TGF-β1-induced epithelial-mesenchymal transition in alveolar epithelial cells through Notch signaling pathway. Toxicol Appl Pharmacol. 318:1–7. 2017. View Article : Google Scholar : PubMed/NCBI
9	Hsu HC, Liu LC, Wang HY, Hung CM, Lin YC, Ho CT and Way TD: Stromal fibroblasts from the interface zone of triple negative breast carcinomas induced epithelial-mesenchymal transition and its inhibition by emodin. PLoS One. 12:e01646612017. View Article : Google Scholar : PubMed/NCBI
10	Li J, Liu P, Mao H, Wanga A and Zhang X: Emodin sensitizes paclitaxel-resistant human ovarian cancer cells to paclitaxel-induced apoptosis in vitro. Oncol Rep. 21:1605–1610. 2009.PubMed/NCBI
11	Ma J, Yang J, Wang C, Zhang N, Dong Y, Wang C, Wang Y and Lin X: Emodin augments cisplatin cytotoxicity in platinum-resistant ovarian cancer cells via ROS-dependent MRP1 downregulation. BioMed Res Int. 2014:1076712014. View Article : Google Scholar : PubMed/NCBI
12	Auersperg N, Wong AS, Choi KC, Kang SK and Leung PC: Ovarian surface epithelium: Biology, endocrinology, and pathology. Endocr Rev. 22:255–288. 2001. View Article : Google Scholar : PubMed/NCBI
13	Alsina-Sanchis E, Figueras A, Lahiguera Á, Vidal A, Casanovas O, Graupera M, Villanueva A and Viñals F: The TGFβ pathway stimulates ovarian cancer cell proliferation by increasing IGF1R levels. Int J Cancer. 139:1894–1903. 2016. View Article : Google Scholar : PubMed/NCBI
14	Lin SW, Lee MT, Ke FC, Lee PP, Huang CJ, Ip MM, Chen L and Hwang JJ: TGFbeta1 stimulates the secretion of matrix metalloproteinase 2 (MMP2) and the invasive behavior in human ovarian cancer cells, which is suppressed by MMP inhibitor BB3103. Clin Exp Metastasis. 18:493–499. 2000. View Article : Google Scholar : PubMed/NCBI
15	Peng H, Liu L and Zhao X: Prognostic significance of matrix metalloproteinase-2 in gynecological cancer: A systemic review of the literature and meta-analysis. J BUON. 18:202–210. 2013.PubMed/NCBI
16	Bartlett JM, Langdon SP, Scott WN, Love SB, Miller EP, Katsaros D, Smyth JF and Miller WR: Transforming growth factor-beta isoform expression in human ovarian tumours. Eur J Cancer. 33:2397–2403. 1997. View Article : Google Scholar : PubMed/NCBI
17	Parikh A, Lee C, Joseph P, Marchini S, Baccarini A, Kolev V, Romualdi C, Fruscio R, Shah H, Wang F, et al: microRNA-181a has a critical role in ovarian cancer progression through the regulation of the epithelial-mesenchymal transition. Nat Commun. 5:29772014. View Article : Google Scholar : PubMed/NCBI
18	Moss EG: MicroRNAs: Hidden in the genome. Curr Biol. 12:R138–R140. 2002. View Article : Google Scholar : PubMed/NCBI
19	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
20	Iorio MV, Visone R, Di Leva G, Donati V, Petrocca F, Casalini P, Taccioli C, Volinia S, Liu CG, Alder H, et al: MicroRNA signatures in human ovarian cancer. Cancer Res. 67:8699–8707. 2007. View Article : Google Scholar : PubMed/NCBI
21	Perdigão-Henriques R, Petrocca F, Altschuler G, Thomas MP, Le MT, Tan SM, Hide W and Lieberman J: miR-200 promotes the mesenchymal to epithelial transition by suppressing multiple members of the Zeb2 and Snail1 transcriptional repressor complexes. Oncogene. 35:158–172. 2016. View Article : Google Scholar : PubMed/NCBI
22	Guo J, Li W, Shi H, Xie X, Li L, Tang H, Wu M, Kong Y, Yang L, Gao J, et al: Synergistic effects of curcumin with emodin against the proliferation and invasion of breast cancer cells through upregulation of miR-34a. Mol Cell Biochem. 382:103–111. 2013. View Article : Google Scholar : PubMed/NCBI
23	Niu G, Li B, Sun L and An C: MicroRNA-153 inhibits osteosarcoma cells proliferation and invasion by targeting TGF-β2. PLoS One. 10:e01192252015. View Article : Google Scholar : PubMed/NCBI
24	Peng T, Zhang S, Li W, Fu S, Luan Y and Zuo L: MicroRNA-141 inhibits glioma cells growth and metastasis by targeting TGF-β2. Am J Transl Res. 8:3513–3521. 2016.PubMed/NCBI
25	Lu R, Ji Z, Li X, Qin J, Cui G, Chen J, Zhai Q, Zhao C, Zhang W and Yu Z: Tumor suppressive microRNA-200a inhibits renal cell carcinoma development by directly targeting TGFB2. Tumour Biol. 36:6691–6700. 2015. View Article : Google Scholar : PubMed/NCBI
26	Zhou Z, Zhang L, Xie B, Wang X, Yang X, Ding N, Zhang J, Liu Q, Tan G, Feng D, et al: FOXC2 promotes chemoresistance in nasopharyngeal carcinomas via induction of epithelial mesenchymal transition. Cancer Lett. 363:137–145. 2015. View Article : Google Scholar : PubMed/NCBI
27	Liu LL, Lu SX, Li M, Li LZ, Fu J, Hu W, Yang YZ, Luo RZ, Zhang CZ and Yun JP: FoxD3-regulated microRNA-137 suppresses tumour growth and metastasis in human hepatocellular carcinoma by targeting AKT2. Oncotarget. 5:5113–5124. 2014. View Article : Google Scholar : PubMed/NCBI
28	He GY, Hu JL, Zhou L, Zhu XH, Xin SN, Zhang D, Lu GF, Liao WT, Ding YQ and Liang L: The FOXD3/miR-214/MED19 axis suppresses tumour growth and metastasis in human colorectal cancer. Br J Cancer. 115:1367–1378. 2016. View Article : Google Scholar : PubMed/NCBI
29	Cheng AS, Li MS, Kang W, Cheng VY, Chou JL, Lau SS, Go MY, Lee CC, Ling TK, Ng EK, et al: Helicobacter pylori causes epigenetic dysregulation of FOXD3 to promote gastric carcinogenesis. Gastroenterology. 144:122–133.e9. 2013. View Article : Google Scholar : PubMed/NCBI
30	Zhang L, Li Z, Gai F and Wang Y: MicroRNA-137 suppresses tumor growth in epithelial ovarian cancer in vitro and in vivo. Mol Med Rep. 12:3107–3114. 2015. View Article : Google Scholar : PubMed/NCBI