Detection of Methylated Septin 9 in Tissue and Plasma of Colorectal Patients with Neoplasia and the Relationship to the Amount of Circulating Cell-free DNA

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Dec 20

PMID 25526039

Citations 62

Authors

Kinga Toth

Reinhold Wasserkort

Ferenc Sipos

Alexandra Kalmar

Barnabas Wichmann

Katalin Leiszter

Gabor Valcz

Mark Juhasz

Pal Miheller

Arpad V Patai

Zsolt Tulassay

Bela Molnar

Affiliations

Soon will be listed here.

Abstract

Background: Determination of methylated Septin 9 (mSEPT9) in plasma has been shown to be a sensitive and specific biomarker for colorectal cancer (CRC). However, the relationship between methylated DNA in plasma and colon tissue of the same subjects has not been reported.

Methods: Plasma and matching biopsy samples were collected from 24 patients with no evidence of disease (NED), 26 patients with adenoma and 34 patients with CRC. Following bisulfite conversion of DNA a commercial RT-PCR assay was used to determine the total amount of DNA in each sample and the fraction of mSEPT9 DNA. The Septin-9 protein was assessed using immunohistochemistry.

Results: The percent of methylated reference (PMR) values for SEPT9 above a PMR threshold of 1% were detected in 4.2% (1/24) of NED, 100% (26/26) of adenoma and 97.1% (33/34) of CRC tissues. PMR differences between NED vs. adenoma and NED vs. CRC comparisons were significant (p<0.001). In matching plasma samples using a PMR cut-off level of 0.01%, SEPT9 methylation was 8.3% (2/24) of NED, 30.8% (8/26) of adenoma and 88.2% (30/34) of CRC. Significant PMR differences were observed between NED vs. CRC (p<0.01) and adenoma vs. CRC (p<0.01). Significant differences (p<0.01) were found in the amount of cfDNA (circulating cell-free DNA) between NED and CRC, and a modest correlation was observed between mSEPT9 concentration and cfDNA of cancer (R2 = 0.48). The level of Septin-9 protein in tissues was inversely correlated to mSEPT9 levels with abundant expression in normals, and diminished expression in adenomas and tumors.

Conclusions: Methylated SEPT9 was detected in all tissue samples. In plasma samples, elevated mSEPT9 values were detected in CRC, but not in adenomas. Tissue levels of mSEPT9 alone are not sufficient to predict mSEPT9 levels in plasma. Additional parameters including the amount of cfDNA in plasma appear to also play a role.

Citing Articles

Advances in blood DNA methylation-based assay for colorectal cancer early detection: a systematic updated review.

Khabbazpour M, Tat M, Karbasi A, Abyazi M, Khodadoustan G, Heidary Z Gastroenterol Hepatol Bed Bench. 2024; 17(3):225-240.

PMID: 39308542 PMC: 11413380. DOI: 10.22037/ghfbb.v17i3.2978.

The common bisulfite-conversion-based techniques to analyze DNA methylation in human cancers.

Jeddi F, Faghfuri E, Mehranfar S, Soozangar N Cancer Cell Int. 2024; 24(1):240.

PMID: 38982390 PMC: 11234524. DOI: 10.1186/s12935-024-03405-2.

DNA methylation modulates epigenetic regulation in colorectal cancer diagnosis, prognosis and precision medicine.

Ye J, Zhang J, Ding W Explor Target Antitumor Ther. 2024; 5(1):34-53.

PMID: 38464391 PMC: 10918240. DOI: 10.37349/etat.2024.00203.

Complete Blood Count Alterations Prior to the Diagnosis of Colorectal Cancer May Help in the Detection of Synchronous Liver Metastases.

Sala R, Ery J, Cuesta-Peredo D, Muedra V, Rodilla V J Clin Med. 2023; 12(20).

PMID: 37892677 PMC: 10607722. DOI: 10.3390/jcm12206540.

Effects of obesity, and of weight loss following bariatric surgery, on methylation of DNA from the rectal mucosa and in cell-free DNA from blood.

Elgendy K, Malcomson F, Afshar S, Bradburn M, Mathers J Int J Obes (Lond). 2023; 47(12):1278-1285.

PMID: 37714902 DOI: 10.1038/s41366-023-01384-4.

References

Lofton-Day C, Model F, deVos T, Tetzner R, Distler J, Schuster M . DNA methylation biomarkers for blood-based colorectal cancer screening. Clin Chem. 2007; 54(2):414-23. DOI: 10.1373/clinchem.2007.095992. View

van der Vaart M, Pretorius P . The origin of circulating free DNA. Clin Chem. 2008; 53(12):2215. DOI: 10.1373/clinchem.2007.092734. View

Levin B, Lieberman D, McFarland B, Andrews K, Brooks D, Bond J . 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. Gastroenterology. 2008; 134(5):1570-95. DOI: 10.1053/j.gastro.2008.02.002. View

Atamaniuk J, Stuhlmeier K, Vidotto C, Tschan H, Dossenbach-Glaninger A, Mueller M . Effects of ultra-marathon on circulating DNA and mRNA expression of pro- and anti-apoptotic genes in mononuclear cells. Eur J Appl Physiol. 2008; 104(4):711-7. DOI: 10.1007/s00421-008-0827-2. View

Grutzmann R, Molnar B, Pilarsky C, Habermann J, Schlag P, Saeger H . Sensitive detection of colorectal cancer in peripheral blood by septin 9 DNA methylation assay. PLoS One. 2008; 3(11):e3759. PMC: 2582436. DOI: 10.1371/journal.pone.0003759. View

Tamkovich S, Cherepanova A, Bryzgunova O, Kolesnikova E, Permyakova V, Vlassov V . Deoxyribonuclease activity in biological fluids of healthy donors and cancer patients. Bull Exp Biol Med. 2009; 146(1):89-91. DOI: 10.1007/s10517-008-0213-4. View

deVos T, Tetzner R, Model F, Weiss G, Schuster M, Distler J . Circulating methylated SEPT9 DNA in plasma is a biomarker for colorectal cancer. Clin Chem. 2009; 55(7):1337-46. DOI: 10.1373/clinchem.2008.115808. View

Lieberman D . Clinical practice. Screening for colorectal cancer. N Engl J Med. 2009; 361(12):1179-87. DOI: 10.1056/NEJMcp0902176. View

Tanzer M, Balluff B, Distler J, Hale K, Leodolter A, Rocken C . Performance of epigenetic markers SEPT9 and ALX4 in plasma for detection of colorectal precancerous lesions. PLoS One. 2010; 5(2):e9061. PMC: 2816214. DOI: 10.1371/journal.pone.0009061. View

10.

Smith R, Cokkinides V, Brooks D, Saslow D, Brawley O . Cancer screening in the United States, 2010: a review of current American Cancer Society guidelines and issues in cancer screening. CA Cancer J Clin. 2010; 60(2):99-119. DOI: 10.3322/caac.20063. View

11.

Amir S, Golan M, Mabjeesh N . Targeted knockdown of SEPT9_v1 inhibits tumor growth and angiogenesis of human prostate cancer cells concomitant with disruption of hypoxia-inducible factor-1 pathway. Mol Cancer Res. 2010; 8(5):643-52. DOI: 10.1158/1541-7786.MCR-09-0497. View

12.

He Q, Chen H, Bai E, Luo Y, Fu R, He Y . Development of a multiplex MethyLight assay for the detection of multigene methylation in human colorectal cancer. Cancer Genet Cytogenet. 2010; 202(1):1-10. DOI: 10.1016/j.cancergencyto.2010.05.018. View

13.

Danese E, Montagnana M, Minicozzi A, De Matteis G, Scudo G, Salvagno G . Real-time polymerase chain reaction quantification of free DNA in serum of patients with polyps and colorectal cancers. Clin Chem Lab Med. 2010; 48(11):1665-8. DOI: 10.1515/CCLM.2010.301. View

14.

Estey M, Kim M, Trimble W . Septins. Curr Biol. 2011; 21(10):R384-7. DOI: 10.1016/j.cub.2011.03.067. View

15.

Toth K, Galamb O, Spisak S, Wichmann B, Sipos F, Valcz G . The influence of methylated septin 9 gene on RNA and protein level in colorectal cancer. Pathol Oncol Res. 2011; 17(3):503-9. DOI: 10.1007/s12253-010-9338-7. View

16.

Connolly D, Yang Z, Castaldi M, Simmons N, Oktay M, Coniglio S . Septin 9 isoform expression, localization and epigenetic changes during human and mouse breast cancer progression. Breast Cancer Res. 2011; 13(4):R76. PMC: 3236340. DOI: 10.1186/bcr2924. View

17.

Ahlquist D, Zou H, Domanico M, Mahoney D, Yab T, Taylor W . Next-generation stool DNA test accurately detects colorectal cancer and large adenomas. Gastroenterology. 2011; 142(2):248-56. PMC: 4017869. DOI: 10.1053/j.gastro.2011.10.031. View

18.

Warren J, Xiong W, Bunker A, Vaughn C, Furtado L, Roberts W . Septin 9 methylated DNA is a sensitive and specific blood test for colorectal cancer. BMC Med. 2011; 9:133. PMC: 3271041. DOI: 10.1186/1741-7015-9-133. View

19.

Ahlquist D, Taylor W, Mahoney D, Zou H, Domanico M, Thibodeau S . The stool DNA test is more accurate than the plasma septin 9 test in detecting colorectal neoplasia. Clin Gastroenterol Hepatol. 2011; 10(3):272-7.e1. PMC: 3980432. DOI: 10.1016/j.cgh.2011.10.008. View

20.

Denters M, Deutekom M, Bossuyt P, Stroobants A, Fockens P, Dekker E . Lower risk of advanced neoplasia among patients with a previous negative result from a fecal test for colorectal cancer. Gastroenterology. 2011; 142(3):497-504. DOI: 10.1053/j.gastro.2011.11.024. View