Advertisement

Tumor Biology

, Volume 36, Issue 2, pp 1245–1250 | Cite as

Preliminary mechanism on the methylation modification of Dkk-1 and Dkk-3 in hepatocellular carcinoma

  • Libo Liang
  • He He
  • Ruixue Lv
  • Mei Zhang
  • Henjian Huang
  • Zhenmei An
  • Shuangqing Li
Research Article

Abstract

Wnt/β-catenin signaling pathway, having a crucial role in regulating diverse cellular processes, can be a new therapeutic target in cancer. To investigate the role of Dkk-1 (Dickkopf-1) and Dkk-3 in tumors and cirrhoses of the liver tissue in hepatocellular carcinoma (HCC), tissues from 38 patients with HCC resections including 5 patients who underwent hemangioma surgery of adjacent tumor tissues at the same time were obtained. Tissues were divided into three groups (nonfibrosis, cirrhosis, and carcinoma) through hematoxylin-eosin (HE) staining. Methylation-specific polymerase chain reaction (PCR) (MSP) measured the methylation status, and reverse transcription-PCR tested the messenger RNA (mRNA) levels, and immunohistochemical analysis provided levels of protein expression. The methylation detection rate of Dkk-1 and Dkk-3 was the highest (P < 0.05) and the mRNA levels of Dkk-1 and Dkk-3 were the lowest (P < 0.05) in the carcinoma tissues. The mRNA levels of β-catenin were significantly higher in the carcinoma tissue than the other tissues (P < 0.05). The expression of Dkk-1 and Dkk-3 was significantly higher in the carcinoma tissues than the other tissues (P < 0.05); but the β-catenin expression was the highest (P < 0.05). Compared with the control, the mRNA levels of β-catenin in the Dkk-1 and Dkk-3 silencing cells increased 5.34 (P < 0.05) and 3.5 times (P > 0.05). After the interference of 5-aza-2′-deoxycytidine, the mRNA levels of Dkk-1 and Dkk-3 significantly increased 58.9 and 59.3 times (P < 0.0001), and the mRNA levels of β-catenin decreased 6.02 times (P < 0.05). In the process of HCC, the abnormal activity of Wnt/β-catenin signaling may be associated with the methylation of Dkk-1 and Dkk-3.

Keywords

Hepatocellular carcinoma Dkk-1 Dkk-3 Methylation 

Notes

Acknowledgments

We thank Ying Binwu for the excellent technical support. This study was supported by West China College of Stomatology Sichuan University and the State Key Laboratory of Biotherapy, West China Hospital of Sichuan University in Chengdu. This study was also supported by the Nature Science Foundation of China (#81100303) and the Science and Technology Department of Sichuan Province (#2014SZ0170).

Authors’ contributions

Liang Libo and He He contributed equally to this work. Liang Libo, He He, and Lv Ruixue designed the study. Liang Libo and Tan Chunlu were involved in the acquisition of data. He He, Zhang Mei, and Huang Hengjian were involved in the interpretation of data. Liang Libo, He He and Li Shuangqing were involved in the manuscript preparation. An Zhenmei provided essential tools.

Conflict of interest

None

Supplementary material

13277_2014_2750_MOESM1_ESM.docx (17 kb)
ESM 1 (DOCX 17 kb)

References

  1. 1.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.CrossRefPubMedGoogle Scholar
  2. 2.
    Chen JG, Zhang SW. Liver cancer epidemic in China: past, present and future. Semin Cancer Biol. 2011;21:59–69.CrossRefPubMedGoogle Scholar
  3. 3.
    Anastas JN, Moon RT. Wnt signalling pathways as therapeutic targets in cancer. Nat Rev Cancer. 2013;13:11–26.CrossRefPubMedGoogle Scholar
  4. 4.
    Ye SL, Chen RX. [comments on management of hepatocellular carcinoma: An update]. Zhonghua gan zang bing za zhi, Zhonghua ganzangbing zazhi, Chin J Hepatol. 2011;19:251–3.Google Scholar
  5. 5.
    Yoda Y, Takeshima H, Niwa T, Kim JG, Ando T, Kushima R, et al. Integrated analysis of cancer-related pathways affected by genetic and epigenetic alterations in gastric cancer. Gastric Canc Offic J Int Gastric Canc Assoc Japan Gastric Canc Assoc. 2014. doi: 10.1007/s10120-014-0348-0.
  6. 6.
    Shen Q, Fan J, Yang XR, Tan Y, Zhao W, Xu Y, et al. Serum dkk1 as a protein biomarker for the diagnosis of hepatocellular carcinoma: a large-scale, multicentre study. Lancet Oncol. 2012;13:817–26.CrossRefPubMedGoogle Scholar
  7. 7.
    Shi J, Keller J, Zhang J, Keller E. A review on the diagnosis and treatment of hepatocellular carcinoma with a focus on the role of wnts and the dickkopf family of wnt inhibitors. J Hepato Carcinom. 2014;1:1–7.Google Scholar
  8. 8.
    Yu B, Yang X, Xu Y, Yao G, Shu H, Lin B, et al. Elevated expression of dkk1 is associated with cytoplasmic/nuclear beta-catenin accumulation and poor prognosis in hepatocellular carcinomas. J Hepatol. 2009;50:948–57.CrossRefPubMedGoogle Scholar
  9. 9.
    Fatima S, Lee NP, Luk JM. Dickkopfs and wnt/beta-catenin signalling in liver cancer. World J Clin Oncol. 2011;2:311–25.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Veeck J, Dahl E. Targeting the wnt pathway in cancer: the emerging role of dickkopf-3. Biochim Biophys Acta. 1825;2012:18–28.Google Scholar
  11. 11.
    Sakaguchi M, Huh NH, Namba M. A novel tumor suppressor, reic/dkk-3 gene identified by our in vitro transformation model of normal human fibroblasts works as a potent therapeutic anti-tumor agent. Adv Exp Med Biol. 2011;720:209–15.CrossRefPubMedGoogle Scholar
  12. 12.
    Lee EJ, Jo M, Rho SB, Park K, Yoo YN, Park J, et al. Dkk3, downregulated in cervical cancer, functions as a negative regulator of beta-catenin. Int J Canc J Int Canc. 2009;124:287–97.CrossRefGoogle Scholar
  13. 13.
    Freese JL, Pino D, Pleasure SJ. Wnt signaling in development and disease. Neurobiol Dis. 2010;38:148–53.CrossRefPubMedGoogle Scholar
  14. 14.
    Baron R, Kneissel M. Wnt signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med. 2013;19:179–92.CrossRefPubMedGoogle Scholar
  15. 15.
    Holland JD, Klaus A, Garratt AN, Birchmeier W. Wnt signaling in stem and cancer stem cells. Curr Opin Cell Biol. 2013;25:254–64.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Libo Liang
    • 1
  • He He
    • 1
  • Ruixue Lv
    • 2
  • Mei Zhang
    • 1
  • Henjian Huang
    • 1
  • Zhenmei An
    • 1
  • Shuangqing Li
    • 1
  1. 1.West China Hospital of Sichuan UniversityChengduChina
  2. 2.MCH Center of GuiyangGuiyangChina

Personalised recommendations