Skip to main content
Download PDF

Disruption of the Tff1 gene in mice using CRISPR/Cas9 promotes body weight reduction and gastric tumorigenesis

Laboratory Animal Research volume 34, pages 257–263 (2018)Cite this article

Abstract

Trefoil factor 1 (TFF1, also known as pS2) is strongly expressed in the gastrointestinal mucosa and plays a critical role in the differentiation of gastric glands. Since approximately 50% of all human gastric cancers are associated with decreased TFF1 expression, it is considered a tumor suppressor gene. Tff1 deficiency in mice results in histological changes in the antral and pyloric gastric mucosa, with severe hyperplasia and dysplasia of epithelial cells, resulting in the development of antropyloric adenoma. Here, we generated Tff1-knockout (KO) mice, without a neomycin resistant (NeoR) cassette, using the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRSIPR/Cas9) system. Though our Tff1-KO mice showed phenotypes very similar to the previous embryonic stem (ES)-cell-based KO mice, they differed from the previous reports in that a reduction in body weight was observed in males. These results demonstrate that these newly established Tff1-KO mice are useful tools for investigating genetic and environmental factors influencing gastric cancer, without the effects of artificial gene insertion. Furthermore, these findings suggest a novel hypothesis that Tff1 expression influences gender differences.

References

  1. Roder DM. The epidemiology of gastric cancer. Gastric Cancer 2002; 5 Suppl 1: 5–11.

    Article  PubMed  Google Scholar 

  2. Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006; 24(14): 2137–2150.

    Article  PubMed  Google Scholar 

  3. Muoz N, Franceschi S. Epidemiology of gastric cancer and perspectives for prevention. Salud Publica Mex 1997; 39(4): 318–330.

    Google Scholar 

  4. Hohenberger P, Gretschel S. Gastric cancer. Lancet 2003; 362(9380): 305–315.

    Article  PubMed  Google Scholar 

  5. Shi SQ, Cai JT, Yang JM. Expression of trefoil factors 1 and 2 in precancerous condition and gastric cancer. World J Gastroenterol 2006; 12(19): 3119–3122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Jakowlew SB, Breathnach R, Jeltsch JM, Masiakowski P, Chambon P. Sequence of the pS2 mRNA induced by estrogen in the human breast cancer cell line MCF-7. Nucleic Acids Res 1984; 12(6): 2861–2878.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Xiao P, Ling H, Lan G, Liu J, Hu H, Yang R. Trefoil factors: Gastrointestinal-specific proteins associated with gastric cancer. Clin Chim Acta 2015; 450: 127–134.

    Article  CAS  PubMed  Google Scholar 

  8. Lefebvre O, Chenard MP, Masson R, Linares J, Dierich A, LeMeur M, Wendling C, Tomasetto C, Chambon P, Rio MC. Gastric mucosa abnormalities and tumorigenesis in mice lacking the pS2 trefoil protein. Science 1996; 274(5285): 259–262.

    Article  CAS  PubMed  Google Scholar 

  9. Ribieras S, Tomasetto C, Rio MC. The pS2/TFF1 trefoil factor, from basic research to clinical applications. Biochim Biophys Acta 1998; 1378(1): F61–77.

    Google Scholar 

  10. Rio MC, Bellocq JP, Daniel JY, Tomasetto C, Lathe R, Chenard MP, Batzenschlager A, Chambon P. Breast cancer-associated pS2 protein: synthesis and secretion by normal stomach mucosa. Science 1988; 241(4866): 705–708.

    Article  CAS  PubMed  Google Scholar 

  11. Wright AV, Nuez JK, Doudna JA. Biology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering. Cell 2016; 164(1-2): 29–44.

    Article  CAS  PubMed  Google Scholar 

  12. Lee JH, Park JH, Nam TW, Seo SM, Kim J Y, Lee HK, Han JH, Park S Y, Choi YK, Lee HW. Differences between immunodeficient mice generated by classical gene targeting and CRISPR/Cas9-mediated gene knockout. Transgenic Res 2018; 27(3): 241–251.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Roh JI, Lee J, Park SU, Kang YS, Lee J, Oh AR, Choi DJ, Cha J Y, Lee HW. CRISPR-Cas9-mediated generation of obese and diabetic mouse models. Exp Anim 2018; 67(2): 229–237.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Sung YH, Kim JM, Kim HT, Lee J, Jeon J, Jin Y, Choi JH, Ban YH, Ha SJ, Kim CH, Lee HW, Kim JS. Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases. Genome Res 2014; 24(1): 125–131.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Henry JA, Bennett MK, Piggott NH, Levett DL, May FE, Westley BR. Expression of the pNR-2/pS2 protein in diverse human epithelial tumours. Br J Cancer 1991; 64(4): 677–682.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Luqmani Y, Bennett C, Paterson I, Corbishley CM, Rio MC, Chambon P, Ryall G. Expression of the pS2 gene in normal, benign and neoplastic human stomach. Int J Cancer 1989; 44(5): 806–812.

    Article  CAS  PubMed  Google Scholar 

  17. Machado JC, Carneiro F, Ribeiro P, Blin N, Sobrinho-Simes M. pS2 protein expression in gastric carcinoma. An immunohistochemical and immunoradiometric study. Eur J Cancer 1996; 32A(9): 1585–1590.

    Article  CAS  PubMed  Google Scholar 

  18. Theisinger B, Welter C, Seitz G, Rio MC, Lathe R, Chambon P, Blin N. Expression of the breast cancer associated gene pS2 and the pancreatic spasmolytic polypeptide gene (hSP) in diffuse type of stomach carcinoma. Eur J Cancer 1991; 27(6): 770–773.

    Article  CAS  PubMed  Google Scholar 

  19. Wanebo HJ, Kennedy BJ, Chmiel J, Steele G Jr, Winchester D, Osteen R. Cancer of the stomach. A patient care study by the American College of Surgeons. Ann Surg 1993; 218(5): 583–592.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Everett SM, Axon AT. Early gastric cancer in Europe. Gut 1997; 41(2): 142–150.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Saukkonen K, Tomasetto C, Narko K, Rio MC, Ristimki A. Cyclooxygenase-2 expression and effect of celecoxib in gastric adenomas of trefoil factor 1-deficient mice. Cancer Res 2003; 63(12): 3032–3036.

    CAS  PubMed  Google Scholar 

  22. Blum D, Stene GB, Solheim TS, Fayers P, Hjermstad MJ, Baracos VE, Fearon K, Strasser F, Kaasa S. Euro-Impact. Validation of the Consensus-Definition for Cancer Cachexia and evaluation of a classification model—a study based on data from an international multicentre project (EPCRC-CSA). Ann Oncol 2014; 25(8): 1635–1642.

    Article  CAS  PubMed  Google Scholar 

  23. Artelt P, Grannemann R, Stocking C, Friel J, Bartsch J, Hauser H. The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene 1991; 99(2): 249–254.

    Article  CAS  PubMed  Google Scholar 

  24. Valera A, Perales JC, Hatzoglou M, Bosch F. Expression of the neomycin-resistance (neo) gene induces alterations in gene expression and metabolism. Hum Gene Ther 1994; 5(4): 449–456.

    Article  CAS  PubMed  Google Scholar 

  25. Alitalo K, Koskinen P, Mkel TP, Saksela K, Sistonen L, Winqvist R. myc oncogenes: activation and amplification. Biochim Biophys Acta 1987; 907(1): 1–32.

    CAS  PubMed  Google Scholar 

Download references

Author information

Author notes

  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, College of Life Science and Biotechnology and Yonsei Laboratory Animal Research Center, Yonsei University, Seoul, 03722, Korea

    Hyejeong Kim, Jaehoon Lee & Han-Woong Lee

  2. Severance Biomedical Science Institute, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea, 03722

    Haengdueng Jeong, Yejin Cho & Ki Taek Nam

Authors
  1. Hyejeong Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haengdueng Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yejin Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jaehoon Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ki Taek Nam
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Han-Woong Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jaehoon Lee, Ki Taek Nam or Han-Woong Lee.

Rights and permissions

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://doi.org/creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kim, H., Jeong, H., Cho, Y. et al. Disruption of the Tff1 gene in mice using CRISPR/Cas9 promotes body weight reduction and gastric tumorigenesis. Lab Anim Res 34, 257–263 (2018). https://doi.org/10.5625/lar.2018.34.4.257

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.5625/lar.2018.34.4.257

Keywords

Laboratory Animal Research

ISSN: 2233-7660