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

  • Original Article
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Pathology & Oncology Research

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.

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References

  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 10:4739–4751

    Article  PubMed  PubMed Central  Google Scholar 

  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 630:1–7

    Article  CAS  PubMed  Google Scholar 

  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 8:84054–84065

    PubMed  PubMed Central  Google Scholar 

  4. Dashwood RH (1999) Early detection and prevention of colorectal cancer (review). Oncol Rep 6:277–281

    CAS  PubMed  Google Scholar 

  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 13:2745–2750

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 8:36354–36367

    PubMed  PubMed Central  Google Scholar 

  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 4:6331

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 6:16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Cahill N, Rosenquist R (2013) Uncovering the DNA methylome in chronic lymphocytic leukemia. Epigenetics 8:138–148

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 8:199

    PubMed  PubMed Central  Google Scholar 

  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 18:263–270

    Article  CAS  PubMed  Google Scholar 

  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 9:126

    Article  PubMed  PubMed Central  Google Scholar 

  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 7:126

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 65:169–177

    CAS  PubMed  Google Scholar 

  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 15:495–501

    Article  PubMed  PubMed Central  Google Scholar 

  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 12:751–763

    Article  PubMed  PubMed Central  Google Scholar 

  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 57:242–245

    Article  PubMed  PubMed Central  Google Scholar 

  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 20:6329–6335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 52:2287–2291

    Article  PubMed  Google Scholar 

  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 64:883–888

    Article  CAS  PubMed  Google Scholar 

  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 190:15–19

    Article  CAS  PubMed  Google Scholar 

  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–S281

    Article  CAS  PubMed  Google Scholar 

  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 7:14265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Yamashita K, Watanabe M (2009) Clinical significance of tumor markers and an emerging perspective on colorectal cancer. Cancer Sci 100:195–199

    Article  CAS  PubMed  Google Scholar 

  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 23:28–33

    Article  PubMed  PubMed Central  Google Scholar 

  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 10:5827–5834

    Article  PubMed  PubMed Central  Google Scholar 

  27. Zarkavelis G, Boussios S, Papadaki A, Katsanos KH, Christodoulou DK, Pentheroudakis G (2017) Current and future biomarkers in colorectal cancer. Ann Gastroenterol 30:613–621

    PubMed  PubMed Central  Google Scholar 

  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 9:55

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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 97:1124–1132

    Article  CAS  PubMed  Google Scholar 

  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 2014:256723

    PubMed  PubMed Central  Google Scholar 

  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 25:3479–3488

    Article  CAS  PubMed  Google Scholar 

  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 42:1920–1933

    Article  CAS  PubMed  Google Scholar 

  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 69:403–8

  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–ee33

    Article  PubMed  Google Scholar 

  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 7:1002–1010

    Article  CAS  Google Scholar 

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Acknowledgments

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

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Authors and Affiliations

  1. Medical Genetics Center, School of Medicine, Ningbo University, Ningbo, Zhejiang, China

    Haochang Hu, Tiangong Wang, Ranran Pan, Yong Yang, Bin Li, Cong Zhou, Jun Zhao & Shiwei Duan

  2. Department of Neurosurgery of Ningbo First Hospital, Ningbo University School of Medicine, Ningbo, Zhejiang, China

    Yi Huang

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  1. Haochang Hu
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  2. Tiangong Wang
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Contributions

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.

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Correspondence to Yi Huang or Shiwei Duan.

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The authors declare no conflicts of interest.

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Haochang Hu and Tiangong Wang are co-first authors of this work.

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Hu, H., Wang, T., Pan, R. et al. Hypermethylated Promoters of Secreted Frizzled-Related Protein Genes are Associated with Colorectal Cancer. Pathol. Oncol. Res. 25, 567–575 (2019). https://doi.org/10.1007/s12253-018-0505-6

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  • DOI: https://doi.org/10.1007/s12253-018-0505-6

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