Serum cell-free DNA concentration in BALB/c mice with azoxymethane-dextran sodium sulfate-induced colorectal cancer

  • Virhan Novianry Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Tanjungpura, Pontianak
  • Yulhasri Yulhasri Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta
  • Kusmardi Kusmardi Department of Pathology Anatomy, Faculty of Medicine, Universitas Indonesia, Jakarta
DOI: https://doi.org/10.13181/mji.v24i1.730
Keywords: azoxymethane, BALB/c mice, cell-free DNA, colorectal cancer, dextran sodium sulfate
Abstract viewed: 1727 times
PDF downloaded: 749 times

Abstract

Background: Colorectal cancer is the third most common cancer in the United States with a mortality rate ranked second in 2012. Early diagnosis such as detection of DNA in serum or faeces at the polyp stage, will reduce colorectal cancer mortality. This study was conducted to analyze the concentration of cell-free DNA (cfDNA) as a tumor marker in colorectal carcinogenesis by using blood serum samples from BALB/c mice previously induced by azoxymethane (AOM) and promoted by dextran sodium sulfate (DSS).

Methods: This experimental animal study used 6 BALB/c mice which had serial intervention in a certain time frame. The first serum samples were taken before induction of carcinogenesis (week-0); then AOM induction of carcinogenesis followed and the second sampling one week after AOM intervention (week-1). Subsequently, promotion of carcinogenesis followed with DSS and the third sampling one week after this intervention (week-2). The fourth sampling was 5 weeks after AOM-DSS intervention (week-6). Quantification of the serum cfDNA was performed with SYBR-Green II fluorescence using Rotor Gene 6000 as a reference. Histopathological examination verified induction of carcinogenesis. For statistical analysis paired T-test was used.

Results: Concentration of serum cfDNA showed significant difference between sampling group at week-0 (1238.49 ± 674.84 pg/μL) and sampling group at week-6 (2244.04 ± 726.57 pg/μL) the latter group showing pre-cancerous histopathology. Slightly increased cfDNA at week-1 with AOM induction (1358.57 ± 803.81 pg/μL) and week-2 after DSS promotion (1317.23 ± 735.92 pg/μL) were not significantly different from week-0 samples.

Conclusion: The concentration of cfDNA in the serum of BALB/c mice 5 weeks after AOM induction of carcinogenesis and DSS promotion is significantly higher than before induction.

Author Biography

Virhan Novianry, Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Tanjungpura, Pontianak
Head of Biochemistry and Molecular Biology Department

References

  1. American Cancer Society. Colorectal cancer facts & figures 2011-2013. Atlanta: American Cancer Society; 2011.

  2. Levin B, Lieberman DA, McFarland B, Smith RA, Brooks D, Andrews KS, et al. Screening and surveillance for the early detection of colorectal cancer and adenomatous polyps, 2008: a joint guideline from the American Cancer Society, the US Multi-Society Task Force on Colorectal Cancer, and the American College of Radiology. CA Cancer J Clin. 2008;58(3):130-60. http://dx.doi.org/10.3322/CA.2007.0018

  3. Reddi KK, Holland JF. Elevated serum ribonuclease in patients with pancreatic cancer. Proc Natl Acad Sci U S A. 1976;73(7):2308-10. http://dx.doi.org/10.1073/pnas.73.7.2308

  4. Kerr DJ, Young AM, Hobbs FDR. ABC of colorectal cCancer. London: BMJ Books; 2001.p.5-7.

  5. Cancer.gov [Internet]. Colon cancer treatment (PDQÃ). [update 2014 Dec 11; cited 2012 Mar 27]. Available from: http://www.cancer.gov/.

  6. Molecular Probes The handbook [Internet]. Nucleic acid detection and quantitation in solution - section 8.3. [cited 2013 Sep 24]. Available from : http//:www.b2b.invitrogen.com/site.

  7. Elshimali YI, Khaddour H, Sarkissyan M, Wu Y, Vadgama JV. The clinical utilization of circulating cell free DNA (CCFDNA) in blood of cancer patients. Int J Mol Sci. 2013. 14(9):18925-58. http://dx.doi.org/10.3390/ijms140918925

  8. Y Lu, L Xie, J Chen. A novel procedure for absolute real-time quantification of gene expression patterns. Plant Methods. 2012;8:1-11. http://dx.doi.org/10.1186/1746-4811-8-9

  9. Wanandi SI, Kurniati V. Sidik DNA (DNA Fingerprinting) dalam biokimia eksperimen laboratorium. Jakarta: Bagian Biokimia FKUI; 2001. p. l-130. Indonesian.

  10. Powell SM, Zilz N, Beazer-Barclay Y, Bryan TM, Hamilton SR, Thibodeau SN, et al. APC mutations occur early during colorectal tumorigenesis. Nature. 1992;359(6392):235-7. http://dx.doi.org/10.1038/359235a0

  11. Gonzalez FJ. CYP2E1. DMD. 2007;35(1):1-8. http://dx.doi.org/10.1124/dmd.106.012492

  12. Chen YQ, Hsieh JT, Yao F, Fang B, Pong RC, Cipriano SC, et al. Induction of apoptosis and G2/M cell cycle arrest by DCC. Oncogene. 1999;18(17):2747-54. http://dx.doi.org/10.1038/sj.onc.1202629

  13. Chen J, Huang FX. The signal pathways in azoxymethane-induced colon cancer and preventive implications. Cancer Biol Ther. 2009;8(14):1313-7. http://dx.doi.org/10.4161/cbt.8.14.8983

  14. Anker P, Stroun M, Maurice PA. Spontaneous release of DNA by human blood lymphocytes as shown in an in vitro system. Cancer Res. 1975;35(9):2375-82.

  15. Leon SA, Shapiro B, Sklaroff DM, Yaros MJ. Free DNA in the serum of cancer patients and the effect of therapy. Cancer Res. 1977;37(3):646-50.

  16. Laroui H, Ingersoll SA, Liu HC, Baker MT, Ayyadurai S, Charania MA, et al. Dextran sodium sulfate (DSS) induces colitis in mice by forming nano-lipocomplexes with medium-chain-length fatty acids in the colon. PLoS One. 2012;7(3):e32084. http://dx.doi.org/10.1371/journal.pone.0032084

  17. N, Triaspolitica. "Mau Nanya Dong Dok." Mau Nanya Dong Dok. N.p, 31 Jan. 1997. Web. 02 July 2017. Retrieved from: https://nanyadongdok.blogspot.com

  18. Czeiger D, Shaked G, Eini H, Vered I, Belochitski O, Avriel A, et al. Measurement of circulating cell-free DNA levels by a new simple fluorescent test in patients with primary colorectal cancer. Am J Clin Pathol. 2011;135(2):264-70. http://dx.doi.org/10.1309/AJCP4RK2IHVKTTZV

Published
2015-03-22
How to Cite
1.
Novianry V, Yulhasri Y, Kusmardi K. Serum cell-free DNA concentration in BALB/c mice with azoxymethane-dextran sodium sulfate-induced colorectal cancer. Med J Indones [Internet]. 2015Mar.22 [cited 2024Apr.20];24(1):3-. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/730
Issue
Vol. 24 No. 1 (2015): March
Section
Basic Medical Research

Copyright (c) 2015 Virhan Novianry, Yulhasri Yulhasri, Kusmardi Kusmardi

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Authors who publish with Medical Journal of Indonesia agree to the following terms:

  1. Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
  2. Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.