Circulating Cell-Free DNA as a Potential Biomarker in Breast Cancer Roles of CA15-3 in breast cancer
Abstract
Background: Breast cancer is the most common cause of death in middle-aged females in western countries. Very high levels of circulating cell-free deoxyribonucleic acid (cfDNA) were found in cancer patients due to contribution of circulating tumor DNA. The aim of this research was to measure circulating cfDNA levels in newly diagnosed breast cancer patients, patients with benign breast disease and healthy controls; investigating its role as a noninvasive biomarker for early detection of breast cancer; and evaluating its sensitivity as a diagnostic marker in breast cancer by comparing its levels with CA15-3 levels.
Methods: This study is a case control study involved 28 patients with primary breast cancer, 15 patients with benign breast tumor, and 10 healthy individuals. The plasma concentration of circulating cfDNA was measured using real‑time PCR.
Results: The study showed a significant difference in the circulating cfDNA levels between cancer group (mean±SD is 77.76±152.76ng/ml) and each of benign group and controls (mean±SD is 0.31±0.87ng/ml and 1.03±1.46ng/ml respectively), with P-value<0.001 for both. There was a statistically significant association between elevated levels of circulating cfDNA and advanced cancer stages with P-value<0.001. Also, circulating cfDNA showed 100% sensitivity in comparison with CA15-3 which showed a sensitivity of 25%.
Conclusion: This study found that plasma levels of cfDNA were considerably higher in cancer patients than in benign group and controls, with cfDNA levels raised significantly when cancer progressed to advanced stages.
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References
Katsura C, Ogunmwonyi I, Kankam HK, Saha S. Breast cancer: presentation, investigation and management. Br J Hosp Med (Lond). 2022;83(2):1-7. doi:10.12968/hmed.2021.0459.
Sun YS, Zhao Z, Yang ZN, et al. Risk Factors and Preventions of Breast Cancer. Int J Biol Sci. 2017;13(11):1387-1397. Published 2017 Nov 1. doi:10.7150/ijbs.21635.
Cardoso F, Harbeck N, Barrios CH, et al. Research needs in breast cancer. Ann Oncol. 2017;28(2):208-217. doi:10.1093/annonc/mdw571.
Buono G, Gerratana L, Bulfoni M, Provinciali N, Basile D, Giuliano M, et al. Circulating tumor DNA analysis in breast cancer: Is it ready for prime-time? Cancer Treatment Reviews. W.B. Saunders Ltd; 2019;73:73–83. doi:10.1016/j.ctrv.2019.01.004.
Cheng F, Su L, Qian C. Circulating tumor DNA: a promising biomarker in the liquid biopsy of cancer. Oncotarget. 2016;7(30):48832-48841. doi:10.18632/oncotarget.9453.
Peled M, Agassi R, Czeiger D, et al. Cell-free DNA concentration in patients with clinical or mammographic suspicion of breast cancer. Sci Rep. 2020;10(1):14601. Published 2020 Sep 3. doi:10.1038/s41598-020-71357-4.
Yu D, Tong Y, Guo X, Feng L, Jiang Z, Ying S, Jia J, et al. Diagnostic Value of Concentration of Circulating Cell-Free DNA in Breast Cancer: A Meta-Analysis. Front Oncol. 2019 Mar 1;9:95. doi: 10.3389/fonc.2019.00095.
Yang J, Cheng L, Zhang J, et al. Predictive value of circulating cell-free DNA in the survival of breast cancer patients: A systemic review and meta-analysis. Medicine (Baltimore). 2018;97(28):e11417. doi:10.1097/MD.0000000000011417.
Mudduwa LK, Wijayaratne GB, Peiris HH, Gunasekera SN, Abeysiriwardhana D, Liyanage N. Elevated pre-surgical CA15-3: does it predict the short-term disease-free survival of breast cancer patients without distant metastasis?. Int J Womens Health. 2018;10:329-335. Published 2018 Jun 26. doi:10.2147/IJWH.S162867.
Alemzadeh E, Allahqoli L, Dehghan H, Mazidimoradi A, Ghasempour A, Salehiniya H. Circulating tumor cells and circulating tumor DNA in breast cancer diagnosis and monitoring. Oncol Res. 2023;31(5):667-675. Published 2023 Jul 21. doi:10.32604/or.2023.028406.
Junqueira-Neto S, Batista IA, Costa JL, Melo SA. Liquid Biopsy beyond Circulating Tumor Cells and Cell-Free DNA. Acta Cytol. 2019;63(6):479-488. doi:10.1159/000493969.
Casadio V, Salvi S. Cell-free DNA as Diagnostic Markers Methods and Protocols Methods in Molecular Biology, New York : Humana Press, 1909, 2019. ISBN 9781493989737
El, Gamaal AS El, Said HH El, Noreldin RI, Omar MMM. Cell-free DNA as a biomarker of breast cancer. Menoufia Med J. 2018;31(2):569. doi:10.4103/mmj.mmj_239_17.
Volik S, Alcaide M, Morin RD, Collins C. Cell-free DNA (cfDNA): Clinical Significance and Utility in Cancer Shaped By Emerging Technologies. Mol Cancer Res. 2016;14(10):898-908. doi:10.1158/1541-7786.MCR-16-0044.
Meddeb R, Dache ZAA, Thezenas S, et al. Quantifying circulating cell-free DNA in humans. Sci Rep. 2019;9(1):5220. doi:10.1038/s41598-019-41593-4.
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):18925-18958. Published 2013 Sep 13. doi:10.3390/ijms140918925.
Fernandez-Garcia D, Hills A, Page K, et al. Plasma cell-free DNA (cfDNA) as a predictive and prognostic marker in patients with metastatic breast cancer. Breast Cancer Res. 2019;21(1):149. Published 2019 Dec 19. doi:10.1186/s13058-019-1235-8.
Atwan, Z.W. GAPDH spike RNA as an alternative for housekeeping genes in relative gene expression assay using real-time PCR. Bull Natl Res Cent (2020);44:32. doi:10.1186/s42269-020-00284-1.
Butera G, Mullappilly N, Masetto F, Palmieri M, Scupoli MT, Pacchiana R, Donadelli M. Regulation of Autophagy by Nuclear GAPDH and Its Aggregates in Cancer and Neurodegenerative Disorders. Int J Mol Sci. 2019 Apr 26;20(9):2062. doi: 10.3390/ijms20092062.
Panagopoulou M, Karaglani M, Balgkouranidou I, et al. Circulating cell-free DNA in breast cancer: size profiling, levels, and methylation patterns lead to prognostic and predictive classifiers. Oncogene. 2019;38(18):3387-3401. doi:10.1038/s41388-018-0660-y.
Bronkhorst AJ, Ungerer V, Holdenrieder S. The emerging role of cell-free DNA as a molecular marker for cancer management. Biomol Detect Quantif. 2019;17:100087. Published 2019 Mar 18. doi:10.1016/j.bdq.2019.100087.
Kustanovich A, Schwartz R, Peretz T, Grinshpun A. Life and death of circulating cell-free DNA. Cancer Biol Ther. 2019;20(8):1057-1067. doi:10.1080/15384047.2019.1598759.
DNA, RNA, and protein purification NucleoSpin Plasma XS kit User Manual. Available from: https://www.bioke.com/webshop/download/18.html?action=download
RealMODTM Green W2 2x qPCR mix method sheet. Available from: https://intronbio.com:6001/inc/downfile.php?filename=F1523866170.pdf&orgfilename=I
Zhang Z, Li L, Shen L, Shen X, Ju S, Cong H. Diagnostic Value of Serum Concentration and Integrity of Circulating Cell-Free DNA in Breast Cancer: A Comparative Study With CEA and CA15-3. Lab Med. 2018;49(4):323-328. doi:10.1093/labmed/lmy019.
Shao Y, Sun X, He Y, Liu C, Liu H. Elevated Levels of Serum Tumor Markers CEA and CA15-3 Are Prognostic Parameters for Different Molecular Subtypes of Breast Cancer. PLoS One. 2015;10(7):e0133830. Published 2015 Jul 24. doi:10.1371/journal.pone.0133830.
Lee JS, Park S, Park JM, Cho JH, Kim SI, Park BW. Elevated levels of preoperative CA15-3 and CEA serum levels have independently poor prognostic significance in breast cancer. Ann Oncol. 2013;24(5):1225-1231. doi:10.1093/annonc/mds604.
Fu Y, Li H. Assessing Clinical Significance of Serum CA15-3 and Carcinoembryonic Antigen (CEA) Levels in Breast Cancer Patients: A Meta-Analysis. Med Sci Monit. 2016;22:3154-3162. Published 2016 Sep 6. doi:10.12659/msm.896563.
Gong B, Xue J, Yu J, et al. Cell-free DNA in blood is a potential diagnostic biomarker of breast cancer. Oncol Lett. 2012;3(4):897-900. doi:10.3892/ol.2012.576.
Hashad D, Sorour A, Ghazal A, Talaat I. Free circulating tumor DNA as a diagnostic marker for breast cancer. J Clin Lab Anal. 2012;26(6):467-472. doi:10.1002/jcla.21548.
Tangvarasittichai O, Jaiwang W, Tangvarasittichai S. The plasma DNA concentration as a potential breast cancer screening marker. Indian J Clin Biochem. 2015;30(1):55-58. doi:10.1007/s12291-013-0407-z
Khurram I, Khan MU, Ibrahim S, Saleem A, Khan Z, Mubeen M, et al. Efficacy of cell-free DNA as a diagnostic biomarker in breast cancer patients. Sci Rep. 2023;13(1):1–9. doi:10.1038/s41598-023-42726-6.
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