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Pathology & Oncology Research

, Volume 25, Issue 2, pp 567–575 | Cite as

Hypermethylated Promoters of Secreted Frizzled-Related Protein Genes are Associated with Colorectal Cancer

  • Haochang Hu
  • Tiangong Wang
  • Ranran Pan
  • Yong Yang
  • Bin Li
  • Cong Zhou
  • Jun Zhao
  • Yi HuangEmail author
  • Shiwei DuanEmail author
Original Article
  • 80 Downloads

Abstract

Colorectal cancer (CRC) is one of the leading causes of death worldwide. Aberrant DNA methylation has been recognized as one of the most common molecular alterations in CRC. The goal of this study was to investigate the diagnostic value of SFRP1 and SFRP2 methylation for CRC. A total of 80 pairs of CRC patients were recruited to test the association of SFRP1 and SFRP2 promotor methylation with CRC. Methylation assay was performed using quantitative methylation-specific polymerase chain reaction (qMSP) method. In this study, we found the methylation levels of SFRP1 and SFRP2 in CRC tumor tissues were significantly higher than those in the adjacent non-tumor tissues (SFRP1: P = 2E-5; SFRP2: P = 0.014). Further bioinformatics analysis of TCGA data confirmed the association of the two genes with CRC (SFRP1: P = 7E-21; SFRP2: P = 5E-24). Luciferase reporter gene assay showed that the recombinant plasmids with SFRP1 and SFRP2 fragments could significantly enhance promoter activity (SFRP1: P = 0.002; SFRP2: P = 0.004). In addition, SFRP1 and SFRP2 methylation were inversely correlated with the mRNA expression displayed by TCGA data mining (SFRP1: r = −0.432, P = 4E-11; SFRP2: r = −0.478, P = 1E-13). GEO data analysis indicated that SFRP1 and SFRP2 expression were increased in three CRC cell lines (COLO320, HCT116 and HT29) after 5′-AZA-deoxycytidine treatment, suggesting that DNA methylation played an important role in regulating gene expression of the two genes. Our results confirmed that promoter methylation of SFRP1 and SFRP2 contributed to the risk of CRC.

Keywords

SFRP1 SFRP2 DNA methylation Colorectal cancer Quantitative methylation specific PCR 

Notes

Acknowledgments

The research is supported by grants from K. C. Wong Magna Fund in Ningbo University.

Author’s Contribution

SD and HH contribute to the conception, design and final approval of the submitted version. TW, CZ, RP and HH contribute to interpretation of data and completion of figures and Tables. YY, BL and JZ contribute to performing the experiments and analyzing the data. HH, TW and SD contribute to writing the paper. All the authors have read and approved the final manuscript.

Compliance with Ethical Standards

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. 1.
    Huang J, Tan ZR, Yu J, Li H, Lv QL, Shao YY, Zhou HH (2017) DNA hypermethylated status and gene expression of PAX1/SOX1 in patients with colorectal carcinoma. Onco Targets Ther 10:4739–4751CrossRefGoogle Scholar
  2. 2.
    Li J, Chen C, Bi X, Zhou C, Huang T, Ni C, Yang P, Chen S, Ye M, Duan S (2017) DNA methylation of CMTM3, SSTR2, and MDFI genes in colorectal cancer. Gene 630:1–7CrossRefGoogle Scholar
  3. 3.
    Hu H, Chen X, Wang C, Jiang Y, Li J, Ying X et al (2017) The role of TFPI2 hypermethylation in the detection of gastric and colorectal cancer. Oncotarget 8:84054–84065Google Scholar
  4. 4.
    Dashwood RH (1999) Early detection and prevention of colorectal cancer (review). Oncol Rep 6:277–281Google Scholar
  5. 5.
    Wu D, Chen X, Xu Y, Wang H, Yu G, Jiang L, Hong Q, Duan S (2017) Prognostic value of MLH1 promoter methylation in male patients with esophageal squamous cell carcinoma. Oncol Lett 13:2745–2750CrossRefGoogle Scholar
  6. 6.
    Liu S, Chen X, Chen R, Wang J, Zhu G, Jiang J et al (2017) Diagnostic role of Wnt pathway gene promoter methylation in non small cell lung cancer. Oncotarget 8:36354–36367Google Scholar
  7. 7.
    Ma D, Jiang C, Hu X, Liu H, Li Q, Li T et al (2014) Methylation patterns of the IFN-gamma gene in cervical cancer tissues. Sci Rep 4:6331CrossRefGoogle Scholar
  8. 8.
    Piras G, Monne M, Palmas AD, Calvisi A, Asproni R, Vacca F, Pilo L, Gabbas A, Latte G (2014) Methylation analysis of the phosphates and tensin homologue on chromosome 10 gene (PTEN) in multiple myeloma. Clin Epigenetics 6:16CrossRefGoogle Scholar
  9. 9.
    Cahill N, Rosenquist R (2013) Uncovering the DNA methylome in chronic lymphocytic leukemia. Epigenetics 8:138–148CrossRefGoogle Scholar
  10. 10.
    Chen C, Wang L, Liao Q, Huang Y, Ye H, Chen F et al (2013) Hypermethylation of EDNRB promoter contributes to the risk of colorectal cancer. Diagn Pathol 8:199Google Scholar
  11. 11.
    Wang Z, Yuan X, Jiao N, Zhu H, Zhang Y, Tong J (2012) CDH13 and FLBN3 gene methylation are associated with poor prognosis in colorectal cancer. Pathol Oncol Res 18:263–270CrossRefGoogle Scholar
  12. 12.
    Oh TJ, Oh HI, Seo YY, Jeong D, Kim C, Kang HW, Han YD, Chung HC, Kim NK, An S (2017) Feasibility of quantifying SDC2 methylation in stool DNA for early detection of colorectal cancer. Clin Epigenetics 9:126CrossRefGoogle Scholar
  13. 13.
    Huang J, Zhang YL, Teng XM, Lin Y, Zheng DL, Yang PY, Han ZG (2007) Down-regulation of SFRP1 as a putative tumor suppressor gene can contribute to human hepatocellular carcinoma. BMC Cancer 7:126CrossRefGoogle Scholar
  14. 14.
    Takeda M, Nagasaka T, Dong-Sheng S, Nishie H, Oka T, Yamada E et al (2011) Expansion of CpG methylation in the SFRP2 promoter region during colorectal tumorigenesis. Acta Med Okayama 65:169–177Google Scholar
  15. 15.
    Park SK, Baek HL, Yu J, Kim JY, Yang HJ, Jung YS, Choi KY, Kim H, Kim HO, Jeong KU, Chun HK, Kim K, Park DI (2017) Is methylation analysis of SFRP2, TFPI2, NDRG4, and BMP3 promoters suitable for colorectal cancer screening in the Korean population? Intest Res 15:495–501CrossRefGoogle Scholar
  16. 16.
    Bartak BK, Kalmar A, Peterfia B, Patai AV, Galamb O, Valcz G et al (2017) Colorectal adenoma and cancer detection based on altered methylation pattern of SFRP1, SFRP2, SDC2, and PRIMA1 in plasma samples. Epigenetics 12:751–763CrossRefGoogle Scholar
  17. 17.
    Babaei H, Mohammadi M, Salehi R (2016) DNA methylation analysis of secreted frizzled-related protein 2 gene for the early detection of colorectal cancer in fecal DNA. Niger Med J 57:242–245CrossRefGoogle Scholar
  18. 18.
    Zhang H, Zhu YQ, Wu YQ, Zhang P, Qi J (2014) Detection of promoter hypermethylation of Wnt antagonist genes in fecal samples for diagnosis of early colorectal cancer. World J Gastroenterol 20:6329–6335CrossRefGoogle Scholar
  19. 19.
    Huang Z, Li L, Wang J (2007) Hypermethylation of SFRP2 as a potential marker for stool-based detection of colorectal cancer and precancerous lesions. Dig Dis Sci 52:2287–2291CrossRefGoogle Scholar
  20. 20.
    Caldwell GM, Jones C, Gensberg K, Jan S, Hardy RG, Byrd P, Chughtai S, Wallis Y, Matthews GM, Morton DG (2004) The Wnt antagonist sFRP1 in colorectal tumorigenesis. Cancer Res 64:883–888CrossRefGoogle Scholar
  21. 21.
    Li B, Chen X, Jiang Y, Yang Y, Zhong J, Zhou C, Hu H, Duan S (2017) CCL2 promoter hypomethylation is associated with gout risk in Chinese Han male population. Immunol Lett 190:15–19CrossRefGoogle Scholar
  22. 22.
    Cannas V, Daino GL, Corona A, Esposito F, Tramontano E (2015) A luciferase reporter gene assay to measure Ebola virus viral protein 35-associated inhibition of double-stranded RNA-stimulated, retinoic acid-inducible gene 1-mediated induction of interferon beta. J Infect Dis 212(Suppl 2):S277–S281CrossRefGoogle Scholar
  23. 23.
    Yu J, Zhai X, Li X, Zhong C, Guo C, Yang F, Yuan Y, Zheng S (2017) Identification of MST1 as a potential early detection biomarker for colorectal cancer through a proteomic approach. Sci Rep 7:14265CrossRefGoogle Scholar
  24. 24.
    Yamashita K, Watanabe M (2009) Clinical significance of tumor markers and an emerging perspective on colorectal cancer. Cancer Sci 100:195–199CrossRefGoogle Scholar
  25. 25.
    Li X, Kong L, Liao S, Lu J, Ma L, Long X (2017) The expression and significance of feces cyclooxygensae-2 mRNA in colorectal cancer and colorectal adenomas. Saudi J Gastroenterol 23:28–33CrossRefGoogle Scholar
  26. 26.
    Wu Y, Gong L, Xu J, Mou Y, Xu X, Qian Z (2017) The clinicopathological significance of HES1 promoter hypomethylation in patients with colorectal cancer. Onco Targets Ther 10:5827–5834CrossRefGoogle Scholar
  27. 27.
    Zarkavelis G, Boussios S, Papadaki A, Katsanos KH, Christodoulou DK, Pentheroudakis G (2017) Current and future biomarkers in colorectal cancer. Ann Gastroenterol 30:613–621Google Scholar
  28. 28.
    Sugai T, Yoshida M, Eizuka M, Uesugii N, Habano W, Otsuka K, Sasaki A, Yamamoto E, Matsumoto T, Suzuki H (2017) Analysis of the DNA methylation level of cancer-related genes in colorectal cancer and the surrounding normal mucosa. Clin Epigenetics 9:55CrossRefGoogle Scholar
  29. 29.
    Chen WD, Han ZJ, Skoletsky J, Olson J, Sah J, Myeroff L, Platzer P, Lu S, Dawson D, Willis J, Pretlow TP, Lutterbaugh J, Kasturi L, Willson JKV, Rao JS, Shuber A, Markowitz SD (2005) Detection in fecal DNA of colon cancer-specific methylation of the nonexpressed vimentin gene. J Natl Cancer Inst 97:1124–1132CrossRefGoogle Scholar
  30. 30.
    Huang S, Zhong X, Gao J, Song R, Wu H, Zi S et al (2014) Coexpression of SFRP1 and WIF1 as a prognostic predictor of favorable outcomes in patients with colorectal carcinoma. Biomed Res Int 2014:256723Google Scholar
  31. 31.
    Veeck J, Niederacher D, An H, Klopocki E, Wiesmann F, Betz B, Galm O, Camara O, Dürst M, Kristiansen G, Huszka C, Knüchel R, Dahl E (2006) Aberrant methylation of the Wnt antagonist SFRP1 in breast cancer is associated with unfavourable prognosis. Oncogene 25:3479–3488CrossRefGoogle Scholar
  32. 32.
    Ba S, Xuan Y, Long ZW, Chen HY, Zheng SS (2017) MicroRNA-27a promotes the proliferation and invasiveness of Colon Cancer cells by targeting SFRP1 through the Wnt/beta-catenin signaling pathway. Cell Physiol Biochem 42:1920–1933CrossRefGoogle Scholar
  33. 33.
    Liu Y, Zhou Q, Zhou D, Huang C, Meng X, Li J (2017) Secreted frizzled-related protein 2-mediated cancer events: Friend or foe? Pharmacol Rep 69:403–8Google Scholar
  34. 34.
    Worm Orntoft MB (2018) Review of blood-based colorectal Cancer screening: how far are circulating cell-free DNA methylation markers from clinical implementation? Clin Colorectal Cancer 17:e415–ee33CrossRefGoogle Scholar
  35. 35.
    Harada T, Yamamoto E, Yamano HO, Nojima M, Maruyama R, Kumegawa K, Ashida M, Yoshikawa K, Kimura T, Harada E, Takagi R, Tanaka Y, Aoki H, Nishizono M, Nakaoka M, Tsuyada A, Niinuma T, Kai M, Shimoda K, Shinomura Y, Sugai T, Imai K, Suzuki H (2014) Analysis of DNA methylation in bowel lavage fluid for detection of colorectal cancer. Cancer Prev Res 7:1002–1010CrossRefGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2018

Authors and Affiliations

  1. 1.Medical Genetics Center, School of MedicineNingbo UniversityNingboChina
  2. 2.Department of Neurosurgery of Ningbo First HospitalNingbo University School of MedicineNingboChina

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