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Levels of plasma circulating cell free nuclear and mitochondrial DNA as potential biomarkers for breast tumors

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Kohler, Corina, Radpour, Ramin, Barekati, Zeinab, Asadollahi, Reza, Bitzer, Johannes, Wight, Edward, Bürki, Nicole, Diesch, Claude, Holzgreve, Wolfgang, Zhong, Xiao Yan·2009·Molecular Cancer
DOI 10.1186/1476-4598-8-105PMC PMC2780981
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Abstract

Background: With the aim to simplify cancer management, cancer research lately dedicated itself more and more to discover and develop non-invasive biomarkers. In this connection, circulating cell-free DNA (ccf DNA) seems to be a promising candidate. Altered levels of ccf nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) have been found in several cancer types and might have a diagnostic value. Methods: Using multiplex real-time PCR we investigated the levels of ccf nDNA and mtDNA in plasma samples from patients with malignant and benign breast tumors, and from healthy controls. To evaluate the applicability of plasma ccf nDNA and mtDNA as a biomarker for distinguishing between the three study-groups we performed ROC (Receiver Operating Characteristic) curve analysis. We also compared the levels of both species in the cancer group with clinicopathological parameters. Results: While the levels of ccf nDNA in the cancer group were significantly higher in comparison with the benign tumor group (P < 0.001) and the healthy control group (P < 0.001), the level of ccf mtDNA was found to be significantly lower in the two tumor-groups (benign: P < 0.001; malignant: P = 0.022). The level of ccf nDNA was also associated with tumor-size (<2 cm vs. >2 cm<5 cm; 2250 vs. 6658; Mann-Whitney-U-Test: P = 0.034). Using ROC curve analysis, we were able to distinguish between the breast cancer cases and the healthy controls using ccf nDNA as marker (cut-off: 1866 GE/ml; sensitivity: 81%; specificity: 69%; P < 0.001) and between the tumor group and the healthy controls using ccf mtDNA as marker (cut-off: 463282 GE/ml; sensitivity: 53%; specificity: 87%; P < 0.001). Conclusion: Our data suggests that nuclear and mitochondrial ccf DNA have potential as biomarkers in breast tumor management. However, ccf nDNA shows greater promise regarding sensitivity and specificity.

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Figure 1 Boxplot for the comparison of the ccf nDNA and mtDNA levels between the study- groups. A) Boxplot for comparison of ccf nDNA levels between the malignant disease group and the healthy control group. Level of ccf nDNA in the cancer group was significantly higher in comparison with the healthy control group (P < 0.001). As no significant difference was found in the level of ccf nDNA between the benign disease group and the healthy controls the comparison is not shown in the figure. B) Boxplot for comparison of ccf mtDNA levels between the tumor group (including the malignant and benign cases) and the healthy control group (P < 0.001). Decreased levels of ccf mtDNA was found in both, the benign disease group and the malignant disease group, when compared to the healthy control group. (* significant correlation; Mann-Whitney-U-Test).
Figure 1 Boxplot for the comparison of the ccf nDNA and mtDNA levels between the study- groups. A) Boxplot for comparison of ccf nDNA levels between the malignant disease group and the healthy control group. Level of ccf nDNA in the cancer group was significantly higher in comparison with the healthy control group (P < 0.001). As no significant difference was found in the level of ccf nDNA between the benign disease group and the healthy controls the comparison is not shown in the figure. B) Boxplot for comparison of ccf mtDNA levels between the tumor group (including the malignant and benign cases) and the healthy control group (P < 0.001). Decreased levels of ccf mtDNA was found in both, the benign disease group and the malignant disease group, when compared to the healthy control group. (* significant correlation; Mann-Whitney-U-Test).CC BY · PubMed Central
Figure 2 Scatterplot for correlating levels of ccf nDNA between breast cancer patients with a tumor size > 5 cm; (n = 4), >2 cm<5 cm; (n = 25) and < 2 cm; (n = 21). Significant difference in the levels of ccf nDNA could be found between tumors with a tumor size of >2 cm<5 cm and tumors with a tumor size of < 2 cm (P = 0.034). For the group of the tumor size > 5 cm, only 4 cases were recruited. (* significant correlation; Mann-Whitney-U-Test).
Figure 2 Scatterplot for correlating levels of ccf nDNA between breast cancer patients with a tumor size > 5 cm; (n = 4), >2 cm<5 cm; (n = 25) and < 2 cm; (n = 21). Significant difference in the levels of ccf nDNA could be found between tumors with a tumor size of >2 cm<5 cm and tumors with a tumor size of < 2 cm (P = 0.034). For the group of the tumor size > 5 cm, only 4 cases were recruited. (* significant correlation; Mann-Whitney-U-Test).CC BY · PubMed Central
Figure 3 ROC curves using ccf nDNA and mtDNA for discriminating between the study-groups. A) ROC curve of ccf nDNA for discriminating between the cancer group and the healthy control group (sensitivity = 81%; specificity = 69%). B) ROC curve of ccf mtDNA for distinguishing between the tumor group and the healthy control group (sensitivity = 53%; specificity = 87%).
Figure 3 ROC curves using ccf nDNA and mtDNA for discriminating between the study-groups. A) ROC curve of ccf nDNA for discriminating between the cancer group and the healthy control group (sensitivity = 81%; specificity = 69%). B) ROC curve of ccf mtDNA for distinguishing between the tumor group and the healthy control group (sensitivity = 53%; specificity = 87%).CC BY · PubMed Central

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Authors
Kohler, C., Radpour, R., Barekati, Z., Asadollahi, R., Bitzer, J., Wight, E., Bürki, N., Diesch, C., Holzgreve, W., & Zhong, X. Y.
Affiliations
  • University of Basel (CH)
  • Kantonsspital Baselland (CH)
  • University Medical Center Freiburg (DE)
Corresponding author
Zhong, Xiao Yan
ORCID
Venue
Molecular Cancer
Publisher
Springer Science and Business Media LLC
Published
2009-11-17
Volume
8
Issue
1
Pages
105-105
ISSN
1476-4598 (ISSN-L)
DOI
10.1186/1476-4598-8-105
PMC
PMC2780981
OpenAlex
W2171143590
Cited by
215
References
39
Citations by year
2026: 3 · 2025: 8 · 2024: 12 · 2023: 15 · 2022: 11 · 2021: 14 · 2020: 15 · 2019: 18 · 2018: 14 · 2017: 11 · 2016: 13 · 2015: 25 · 2014: 9 · 2013: 15 · 2012: 11
FWCI
3.46
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Molecular Cancer (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • National Science Foundation
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (320000, 119722)
  • Krebsliga Beider Basel
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
  • Health Sciences › Medicine › Pediatrics, Perinatology and Child Health › Prenatal Screening and Diagnostics
Keywords
cell-free DNA, cell-free mitochondrial DNA, breast cancer, benign breast tumors, plasma biomarkers, tumor size, sensitivity and specificity
MeSH terms
Breast Neoplasms, Cell Nucleus, Cell-Free System, Diagnosis, Differential, DNA, Mitochondrial, Female, Health, Humans, Lymph Nodes, Neoplasm Metastasis, Receptors, Estrogen, Receptors, Progesterone, ROC Curve, Biomarkers, Tumor, Cohort Studies, Case-Control Studies, Erb-b2 Receptor Tyrosine Kinases

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