Effects of emodin on the demethylation of tumor-suppressor genes in pancreatic cancer PANC-1 cells

Zhang,Hao; Chen,Liang; Bu,He-Qi; Yu,Qing-Jiang; Jiang,Dan-Dan; Pan,Feng-Ping; Wang,Yu; Liu,Dian-Lei; Lin,Sheng-Zhang

doi:10.3892/or.2015.3914

Effects of emodin on the demethylation of tumor-suppressor genes in pancreatic cancer PANC-1 cells

Authors:
- Hao Zhang
- Liang Chen
- He-Qi Bu
- Qing-Jiang Yu
- Dan-Dan Jiang
- Feng-Ping Pan
- Yu Wang
- Dian-Lei Liu
- Sheng-Zhang Lin
View Affiliations

Affiliations: Department of Hepatobiliary Surgery, The First Hospital of Jiaxing, Jiaxing, Zhejiang 310012, P.R. China, Department of Anorectal Surgery, Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang 310012, P.R. China, Department of Surgery, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang 310003, P.R. China, Department of Hepatobiliary-Pancreatic Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003, P.R. China
Published online on: April 17, 2015 https://doi.org/10.3892/or.2015.3914
Pages: 3015-3023

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

Emodin, a natural anthraquinone derivative isolated from Rheum palmatum, has been reported to inhibit the growth of pancreatic cancer cells through different modes of action; yet, the detailed mechanism remains unclear. In the present study, we hypothesized that emodin exerts its antitumor effect by participating in the regulation of the DNA methylation level. Our research showed that emodin inhibited the growth of pancreatic cancer PANC-1 cells in a dose- and time-dependent manner. Dot-blot results showed that 40 µM emodin significantly inhibited genomic 5mC expression in the PANC-1 cells, and mRNA‑Seq showed that different concentrations of emodin could alter the gene expression profile in the PANC-1 cells. BSP confirmed that the methylation levels of P16, RASSF1A and ppENK were decreased, while concomitantly the unmethylated status was increased. RT-PCR and western blotting results confirmed that the low expression or absence of expression of mRNA and protein in the PANC-1 cells was re-expressed following treatment with emodin. In conclusion, our study for the first time suggests that emodin inhibits pancreatic cancer cell growth, which may be related to the demethylation of tumor-suppressor genes. The related mechanism may be through the inhibition of methyltransferase expression.

View Figures

View References

1	Reske SN: PET and PET-CT of malignant tumors of the exocrine pancreas. Radiologe. 49:131–136. 2009.In German. View Article : Google Scholar : PubMed/NCBI
2	Hariharan D, Saied A and Kocher HM: Analysis of mortality rates for pancreatic cancer across the world. HPB Oxf. 10:58–62. 2008. View Article : Google Scholar
3	Guo X and Cui Z: Current diagnosis and treatment of pancreatic cancer in China. Pancreas. 31:13–22. 2005. View Article : Google Scholar : PubMed/NCBI
4	Cheng X and Blumenthal RM: Mammalian DNA methyltransferases: a structural perspective. Structure. 16:341–350. 2008. View Article : Google Scholar : PubMed/NCBI
5	Bird A: DNA methylation patterns and epigenetic memory. Genes Dev. 16:6–21. 2002. View Article : Google Scholar : PubMed/NCBI
6	Gambichler T, Sand M and Skrygan M: Loss of 5-hydroxymeth-ylcytosine and ten-eleven translocation 2 protein expression in malignant melanoma. Melanoma Res. 23:218–220. 2013. View Article : Google Scholar : PubMed/NCBI
7	Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, Harper JW and Elledge SJ: p57^KIP2, a structurally distinct member of the p21^CIP1 Cdk inhibitor family, is a candidate tumor suppressor gene. Genes Dev. 9:650–662. 1995. View Article : Google Scholar : PubMed/NCBI
8	Sato N, Fukushima N, Maehara N, Matsubayashi H, Koopmann J, Su GH, Hruban RH and Goggins M: SPARC/osteonectin is a frequent target for aberrant methylation in pancreatic adenocarcinoma and a mediator of tumor-stromal interactions. Oncogene. 22:5021–5030. 2003. View Article : Google Scholar : PubMed/NCBI
9	Attri J, Srinivasan R, Majumdar S, Radotra BD and Wig J: Alterations of tumor suppressor gene P16INK4a in pancreatic ductal carcinoma. BMC Gastroenterol. 5:222005. View Article : Google Scholar : PubMed/NCBI
10	Dammann R, Schagdarsurengin U, Liu L, Otto N, Gimm O, Dralle H, Boehm BO, Pfeifer GP and Hoang-Vu C: Frequent RASSF1A promoter hypermethylation and K-ras mutations in pancreatic carcinoma. Oncogene. 22:3806–3812. 2003. View Article : Google Scholar : PubMed/NCBI
11	Ueki T, Toyota M, Skinner H, Walter KM, Yeo CJ, Issa JP, Hruban RH and Goggins M: Identification and characterization of differentially methylated CpG islands in pancreatic carcinoma. Cancer Res. 61:8540–8546. 2001.PubMed/NCBI
12	Fukushima N, Sato N, Ueki T, Rosty C, Walter KM, Wilentz RE, Yeo CJ, Hruban RH and Goggins M: Aberrant methylation of preproenkephalin and P16 genes in pancreatic intraepithelial neoplasia and pancreatic ductal adenocarcinoma. Am J Pathol. 160:1573–1581. 2002. View Article : Google Scholar : PubMed/NCBI
13	Schutte M, Hruban RH, Geradts J, Maynard R, Hilgers W, Rabindran SK, Moskaluk CA, Hahn SA, Schwarte-Waldhoff I, Schmiegel W, et al: Abrogation of the Rb/P16 tumor-suppressive pathway in virtually all pancreatic carcinomas. Cancer Res. 57:3126–3130. 1997.PubMed/NCBI
14	Moore PS, Sipos B, Orlandini S, Sorio C, Real FX, Lemoine NR, Gress T, Bassi C, Klöppel G, Kalthoff H, et al: Genetic profile of 22 pancreatic carcinoma cell lines. Analysis of K-ras, p53, P16 and DPC4/Smad4. Virchows Arch. 439:798–802. 2001. View Article : Google Scholar
15	Brueckner B, Kuck D and Lyko F: DNA methyltransferase inhibitors for cancer therapy. Cancer J. 13:17–22. 2007. View Article : Google Scholar : PubMed/NCBI
16	Szyf M: DNA methylation and demethylation as targets for anticancer therapy. Biochemistry (Mosc). 70:533–549. 2005. View Article : Google Scholar
17	Goffin J and Eisenhauer E: DNA methyltransferase inhibitors - state of the art. Ann Oncol. 13:1699–1716. 2002. View Article : Google Scholar : PubMed/NCBI
18	Lyko F and Brown R: DNA methyltransferase inhibitors and the development of epigenetic cancer therapies. J Natl Cancer Inst. 97:1498–1506. 2005. View Article : Google Scholar : PubMed/NCBI
19	Xiang N, Zhao R, Song G and Zhong W: Selenite reactivates silenced genes by modifying DNA methylation and histones in prostate cancer cells. Carcinogenesis. 29:2175–2181. 2008. View Article : Google Scholar : PubMed/NCBI
20	Brueckner B, Garcia Boy R, Siedlecki P, Musch T, Kliem HC, Zielenkiewicz P, Suhai S, Wiessler M and Lyko F: Epigenetic reactivation of tumor suppressor genes by a novel small-molecule inhibitor of human DNA methyltransferases. Cancer Res. 65:6305–6311. 2005. View Article : Google Scholar : PubMed/NCBI
21	Cui X, Wakai T, Shirai Y, Yokoyama N, Hatakeyama K and Hirano S: Arsenic trioxide inhibits DNA methyltransferase and restores methylation-silenced genes in human liver cancer cells. Hum Pathol. 37:298–311. 2006. View Article : Google Scholar : PubMed/NCBI
22	Lin SZ, Chen KJ, Tong HF, Jing H, Li H and Zheng SS: Emodin attenuates acute rejection of liver allografts by inhibiting hepatocellular apoptosis and modulating the Th1/Th2 balance in rats. Clin Exp Pharmacol Physiol. 37:790–794. 2010.PubMed/NCBI
23	Li HL, Chen HL, Li H, Zhang KL, Chen XY, Wang XW, Kong QY and Liu J: Regulatory effects of emodin on NF-κB activation and inflammatory cytokine expression in RAW 264.7 macrophages. Int J Mol Med. 16:41–47. 2005. View Article : Google Scholar : PubMed/NCBI
24	Li-Weber M: Targeting apoptosis pathways in cancer by Chinese medicine. Cancer Lett. 332:304–312. 2013. View Article : Google Scholar
25	Liu A, Chen H, Tong H, Ye S, Qiu M, Wang Z, Tan W, Liu J and Lin S: Emodin potentiates the antitumor effects of gemcitabine in pancreatic cancer cells via inhibition of nuclear factor-κB. Mol Med Rep. 4:221–227. 2011.PubMed/NCBI
26	Liu Z, Xie Z, Jones W, Pavlovicz RE, Liu S, Yu J, Li PK, Lin J, Fuchs JR, Marcucci G, et al: Curcumin is a potent DNA hypomethylation agent. Bioorg Med Chem Lett. 19:706–709. 2009. View Article : Google Scholar
27	Fang MZ, Wang Y, Ai N, Hou Z, Sun Y, Lu H, Welsh W and Yang CS: Tea polyphenol (−)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res. 63:7563–7570. 2003.PubMed/NCBI
28	Xu W, Yang H, Liu Y, Yang Y, Wang P, Kim SH, Ito S, Yang C, Wang P, Xiao MT, et al: Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxy-genases. Cancer Cell. 19:17–30. 2011. View Article : Google Scholar : PubMed/NCBI
29	Lin SZ, Wei WT, Chen H, Chen KJ, Tong HF, Wang ZH, Ni ZL, Liu HB, Guo HC and Liu DL: Antitumor activity of emodin against pancreatic cancer depends on its dual role: promotion of apoptosis and suppression of angiogenesis. PLoS One. 7:e421462012. View Article : Google Scholar : PubMed/NCBI
30	Liu A, Luo J and Zhang JH: Emodin combined gemcitabine inhibited the growth of pancreatic cancer in vitro and in vivo and its mechanisms study. Zhongguo Zhong Xi Yi Jie He Za Zhi. 32:652–656. 2012.In Chinese. PubMed/NCBI
31	Chen H, Wei W, Guo Y, Liu A, Tong H, Wang Z, Tan W, Liu J and Lin S: Enhanced effect of gemcitabine by emodin against pancreatic cancer in vivo via cytochrome C-regulated apoptosis. Oncol Rep. 25:1253–1261. 2011.PubMed/NCBI
32	Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y, Agarwal S, Iyer LM, Liu DR, Aravind L, et al: Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science. 324:930–935. 2009. View Article : Google Scholar : PubMed/NCBI
33	Wu SC and Zhang Y: Active DNA demethylation: many roads lead to Rome. Nat Rev Mol Cell Biol. 11:607–620. 2010. View Article : Google Scholar : PubMed/NCBI
34	Yang H, Liu Y, Bai F, Zhang JY, Ma SH, Liu J, Xu ZD, Zhu HG, Ling ZQ, Ye D, et al: Tumor development is associated with decrease of TET gene expression and 5-methylcytosine hydroxylation. Oncogene. 32:663–669. 2013. View Article : Google Scholar