Clinical relevance of circulating biomarkers in breast cancer metastasis detection: "insights from liquid biopsy technology"

Zohreh Salimi , Rasoul Fatehifard, Mahmoud Aghaei, Mojtaba Panjehpour

Abstract


Background and purpose: Breast cancer is a major global concern, especially because of high mortality linked to advanced stages. Metastasis may occur early, but current diagnostics struggle to detect small metastatic cells. Non-invasive liquid biopsies, such as circulating miRNA, ctDNA, and cfDNA, present a promising solution for screening and monitoring breast cancer. This study aimed to examine the clinical relevance of circulating levels of cfDNA, miR-182, CYFRA21-1, and CEA in metastatic and non-metastatic breast cancer patients and their association with clinical stage and distant metastasis.

Experimental approach: Ten mL blood samples were collected from 17 metastatic and 29 non-metastatic breast cancer patients pre-surgery. cfDNA was measured fluorometrically, miRNA-182 via quantitative real-time PCR, CYFRA21-1, and CEA using enzyme-linked immunosorbent assay ELISA.

Findings/Results: Results demonstrated significantly higher levels of cfDNA, miR-182, CYFRA21-1, and CEA in women with metastatic breast cancer compared to those with non-metastatic breast cancer. These markers were linked to advanced clinical stages and increased tumor size. Elevated levels of cfDNA, miR-182, and CYFRA21-1 were indicative of a higher risk for lymphatic metastasis, while cfDNA and CYFRA21-1 were associated with distant metastasis. ROC curve analyses revealed strong efficacy for cfDNA (AUC 0.94), miR-182 (AUC 0.91), CYFRA21-1 (AUC 0.88), and CEA (AUC 0.87) in detecting metastatic breast cancer.

Conclusion and implications: Combined analysis of these biomarkers will enhance the predictive accuracy of metastatic breast cancer and clarify the relationship between biomarker profiles and the characteristics of metastatic versus nonmetastatic patients using liquid biopsy technology.

 

 


Keywords


Biomarkers; cfDNA; CYFRA21-1; Liquid biopsy; Metastatic breast cancer; miR-182.

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References


Sariego J. Breast cancer in the young patient. Am Surg. 2010;76(12):1397-1400.PMID: 21265355.

Cardoso F, Senkus E, Costa A, Papadopoulos E, Aapro M, André F, et al. 4th ESO–ESMO international consensus guidelines for advanced breast cancer (ABC 4). Ann Oncol. 2018;29(8):1634-1657.DOI: 10.1093/annonc/mdy192.

O'Shaughnessy J. Extending survival with chemotherapy in metastatic breast cancer. Oncologist. 2005;10(S3):20-29.DOI: 10.1634/theoncologist.10-90003-20.

Vogsen M, Harbo F, Jakobsen NM, Nissen HJ, Dahlsgaard-Wallenius SE, Gerke O, et al. Response monitoring in metastatic breast Cancer:a prospective study comparing 18F-FDG PET/CT with conventional CT. J Nucl Med. 2023;64(3):355-361.DOI: 10.2967/jnumed.121.263358.

Zhu JW, Charkhchi P, Akbari MR. Potential clinical utility of liquid biopsies in ovarian cancer. Mol Cancer. 2022;21(1):114,1-24.DOI: 10.1186/s12943-022-1588-8.

Dawson SJ, Tsui DW, Murtaza M, Biggs H, Rueda OM, Chin SF, et al. Analysis of circulating tumor DNA to monitor metastatic breast cancer. N Engl J Med. 2013;368(13):1199-1209.DOI: 10.1056/NEJMoa1213261.

Segura MF, Hanniford D, Menendez S, Reavie L, Zou X, Alvarez-Diaz S, et al. Aberrant miR-182 expression promotes melanoma metastasis by repressing FOXO3 and microphthalmia-associated transcription factor. Proc Natl Acad Sci U S A. 2009;106(6):1814-1819.DOI: 10.1073/pnas.0808263106.

Soltani S, Mokarian F, Panjehpour M. The expression of CK-19 gene in circulating tumor cells of blood samples of metastatic breast cancer women. Res Pharm Sci. 2015;10(6):485-496.PMID: 26779268.

Marrakchi R, Ouerhani S, Benammar S, Rouissi K, Bouhaha R, Bougatef K, et al. Detection of cytokeratin 19 mRNA and Cyfra 21–1 (cytokeratin 19 fragments) in blood of Tunisian women with breast cancer. Int J Biol Markers. 2008;23(4):238-243.DOI: 10.1177/172460080802300407.

Sundström B, Stigbrand T. Cytokeratins and tissue polypeptide antigen. Int J Biol Markers. 1994;9(2):102-108.DOI: 10.1177/172460089400900207.

Fernandez-Gomez J, Rodríguez-Martínez JJ, Barmadah SE, Rodríguez JG, Allende M, Jalon A, et al. Urinary CYFRA 21.1 is not a useful marker for the detection of recurrences in the follow-up of superficial bladder cancer. Eur Urol. 2007;51(5):1267-1274.DOI: 10.1016/j.eururo.2006.12.019.

Stieber P, Dienemann H, Hasholzner U, Fabricius P, Schambeck C, Weinzierl M, et al. Comparison of CYFRA 21-1, TPA and TPS in lung cancer, urinary bladder cancer and benign diseases. Int J Biol Markers. 1994;9(2):82-88.DOI: 10.1177/172460089400900204.

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):e0133830,1-11.DOI: 10.1371/journal.pone.0133830.

Zhu W, Zhou K, Zha Y, Chen D, He J, Ma H, et al. Diagnostic value of serum miR-182, miR-183, miR-210, and miR-126 levels in patients with early-stage non-small cell lung cancer. PloS One. 2016;11(4):e0153046.DOI: 10.1371/journal.pone.0153046.

Hanafi AR, Jayusman AM, Imelda P, Alfasunu S, Sadewa AH, Pramono D, et al. CEA and Cyfra 21-1 linked to serial miRNA expressions of advanced-stage non-small cell lung cancer in Indonesia. J Med Sci. 2023;55(4):316-325.DOI: 10.19106/JMedSci005504202303.

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):264-270.DOI: 10.1309/AJCP4RK2IHVKTTZV.

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):15347,1-9. DOI: 10.1038/s41598-023-42726-6.

Sopik V, Narod SA. The relationship between tumour size, nodal status and distant metastases: on the origins of breast cancer. Breast Cancer Res Treat. 2018;170(2):647-656.DOI: 10.1007/s10549-018-4796-9.

Chen R, Qarmali M, Siegal GP, Wei S. Receptor conversion in metastatic breast cancer: analysis of 390 cases from a single institution. Mod Pathol. 2020;33(12):2499-2506.DOI: 10.1038/s41379-020-0615-z.

Haghshenas M, Firouzabadi N, Akbarizadeh AR, Rashedinia M. Combination of metformin and gallic acid induces autophagy and apoptosis in human breast cancer cells. Res Pharm Sci . 2023;18(6):663-675.DOI: 10.4103/1735-5362.389956.

Wang PY, Gong HT, Li BF, Lv CL, Wang HT, Zhou HH, et al. Higher expression of circulating miR‑182 as a novel biomarker for breast cancer. Oncol Lett. 2013;6(6):1681-1886.DOI: 10.3892/ol.2013.1593.

Nakata B, Takashima T, Ogawa Y, Ishikawa T, Hirakawa K. Serum CYFRA 21-1 (cytokeratin-19 fragments) is a useful tumour marker for detecting disease relapse and assessing treatment efficacy in breast cancer. Br J Cancer. 2004;91(5):873-878.DOI: 10.1038/sj.bjc.6602074.

Kabel AM. Tumor markers of breast cancer: new prospectives. J Oncol. 2017;3(1):5-11.DOI: 10.1016/j.jons.2017.01.001.

Shreffler J, Huecker MR. Diagnostic testing accuracy: sensitivity, specificity, predictive values and likelihood ratios. Last updated 2023. In: StatPearls. Treasure Island (FL). StatPearls Publishing; 2025. Available at: https://www.ncbi.nlm.nih.gov/books/NBK557491/PMID: 32491423.

Coombes RC, Page K, Salari R, Hastings RK, Armstrong A, Ahmed S, et al. Personalized detection of circulating tumor DNA antedates breast cancer metastatic recurrence. Clin Cancer Res. 2019;25(14):4255-4263.DOI: 10.1158/1078-0432.CCR-18-3663.

Zhong Y, Fan Q, Zhou Z, Wang Y, He K, Lu J. Plasma cfDNA as a potential biomarker to evaluate the efficacy of chemotherapy in gastric cancer. Cancer Manag. Res. 2020;12:3099-3106.DOI: 10.2147/CMAR.S243320.

Pu WY, Zhang R, Xiao L, Wu YY, Gong W, Lv XD, et al. Prediction of cancer progression in a group of 73 gastric cancer patients by circulating cell-free DNA. BMC Cancer. 2016;16(1):943,1-5.DOI: 10.1186/s12885-016-2977-7.

Salvianti F, Giuliani C, Petrone L, Mancini I, Vezzosi V, Pupilli C, et al. Integrity and quantity of total cell-free DNA in the diagnosis of thyroid cancer: correlation with cytological classification. Int J Mol Sci. 2017;18(7):1350,1-10.DOI: 10.3390/ijms18071350.

Shao X, He Y, Ji M, Chen X, Qi J, Shi W, et al. Quantitative analysis of cell-free DNA in ovarian cancer. Oncol Lett. 2015;10(6):3478-3482.DOI: 10.3892/ol.2015.3771.

Fettke H, Kwan EM, Bukczynska P, Ng N, Nguyen-Dumont T, Southey MC, et al. Prognostic impact of total plasma cell-free DNA concentration in androgen receptor pathway inhibitor–treated metastatic castration-resistant prostate cancer. Eur Urol Focus. 2021;7(6):1287-1291.DOI: 10.1016/j.euf.2020.07.001.

Nie K, Jia Y, Zhang X. Cell-free circulating tumor DNA in plasma/serum of non-small cell lung cancer. Tumour Biol. 2015;36(1):7-19.DOI: 10.1007/s13277-014-2758-3.

Singh N, Gupta S, Pandey R, Chauhan SS, Saraya A. High levels of cell-free circulating nucleic acids in pancreatic cancer are associated with vascular encasement, metastasis and poor survival. Cancer Investig. 2015;33(3):78-85.DOI: 10.3109/07357907.2014.1001894

Schaefer A, Jung M, Mollenkopf HJ, Wagner I, Stephan C, Jentzmik F, et al. Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma. Int J Cancer. 2010;126(5):1166-1176.DOI: 10.1002/ijc.24827.

Han Y, Chen J, Zhao X, Liang C, Wang Y, Sun L, et al. MicroRNA expression signatures of bladder cancer revealed by deep sequencing. PLoS One. 2011;6(3):e18286,1-6.DOI: 10.1371/journal.pone.0018286.

Yamada Y, Enokida H, Kojima S, Kawakami K, Chiyomaru T, Tatarano S, et al. MiR‐96 and miR‐183 detection in urine serve as potential tumor markers of urothelial carcinoma: correlation with stage and grade, and comparison with urinary cytology. Cancer Sci. 2011;102(3):522-529.DOI: 10.1111/j.1349-7006.2010.01816.x.

Mahmoud HA, El Amin HA, Ahmed ESM, Kenawy AG, El-Ebidi AM, ElNakeeb I, et al. Role of MicroRNA-223 and MicroRNA-182 as novel biomarkers in early detection of colorectal cancer. Int J Gen Med. 2022,15:3281-3291.DOI: 10.2147/IJGM.S353244.

Ali OS, Shabayek MI, Seleem MM, Abdelaziz HG, Makhlouf DO. MicroRNAs 182 and 375 sera expression as prognostic biochemical markers in breast cancer. Clin Breast Cancer. 2018;18(6):e1373-e1379.DOI: 10.1016/j.clbc.2018.07.020.

Abramovic I, Vrhovec B, Skara L, Vrtaric A, Nikolac Gabaj N, Kulis T, et al. MiR-182-5p and miR-375-3p have higher performance than PSA in discriminating prostate cancer from benign prostate hyperplasia. Cancers. 2021;13(9): 2068,1-13.DOI: 10.3390/cancers13092068.

Ebied SA, Abdel-Rehim WM, El-Benhawy SA, El-Gawish MA, Hassan MA, El-Settawy II. Serum CYFRA 21-1 in Egyptian women with breast cancer. Alex J Med. 2017;53(1):41-47.DOI: 10.1016/j.ajme.2016.02.006.

Bidard FC, Hajage D, Bachelot T, Delaloge S, Brain E, Campone M, et al. Assessment of circulating tumor cells and serum markers for progression-free survival prediction in metastatic breast cancer:a prospective observational study. Breast Cancer Res. 2012;14(1):R29,1-10.DOI: 10.1186/bcr3114.

Giovanella L, Imperiali M, Trimboli P. Role of serum cytokeratin 19 fragment (Cyfra 21.1) as a prognostic biomarker in patients with differentiated thyroid cancer. Sci Rep. 2017;7(1):7359,1-7.DOI: 10.1038/s41598-017-07915-0.

Li J, Chen Y, Wang X, Wang C, Xiao M. The value of combined detection of CEA, CYFRA21-1, SCC-Ag, and pro-GRP in the differential diagnosis of lung cancer. Transl Cancer Res. 2021;10(4):1900-1906.DOI: 10.21037/tcr-21-527.

He ZY, Li X, Chen QS, Sun JY, Li FY, Wu SG, et al. Elevated serum carcinoembryonic antigen and CA15-3 levels and the risk of site-specific metastases in metastatic breast cancer. Transl Cancer Res. 2016;5(5):529-537.DOI: 10.21037/tcr.2016.08.39.

Nam Se, Lim W, Jeong J, Lee S, Choi J, Park H. The prognostic significance of preoperative tumor marker (CEA, CA15-3) elevation in breast cancer patients: data from the Korean Breast Cancer Society Registry. Breast Cancer Res Treat. 2019;177(3):669-678.DOI: 10.1007/s10549-019-05357-y.

Hao C, Zhang G, Zhang L. Serum CEA levels in 49 different types of cancer and noncancer diseases. Progr Mol Biol Transl Sci. 2019;162:213-227.DOI: 10.1016/bs.pmbts.2018.12.011.


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