Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells
The true importance of cell-free DNA in human biology, together with the potential scale of its clinical utility, is tarnished by a lack of understanding of its composition and origin. In investigating the cell-free DNA present in the growth medium of cultured 143B cells, we previously demonstrated...
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SAGE Publishing
2018-09-01
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| Series: | Tumor Biology |
| Online Access: | https://doi.org/10.1177/1010428318801190 |
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| author | Abel Jacobus Bronkhorst Johannes F Wentzel Vida Ungerer Dimetrie L Peters Janine Aucamp Etienne Pierre de Villiers Stefan Holdenrieder Piet J Pretorius |
| author_facet | Abel Jacobus Bronkhorst Johannes F Wentzel Vida Ungerer Dimetrie L Peters Janine Aucamp Etienne Pierre de Villiers Stefan Holdenrieder Piet J Pretorius |
| author_sort | Abel Jacobus Bronkhorst |
| collection | DOAJ |
| description | The true importance of cell-free DNA in human biology, together with the potential scale of its clinical utility, is tarnished by a lack of understanding of its composition and origin. In investigating the cell-free DNA present in the growth medium of cultured 143B cells, we previously demonstrated that the majority of cell-free DNA is neither a product of apoptosis nor necrosis. In the present study, we investigated the composition and origin of this cell-free DNA population using next-generation sequencing. We found that the cell-free DNA comprises mainly of repetitive DNA, including α-satellite DNA, mini satellites, and transposons that are currently active or exhibit the capacity to become reactivated. A significant portion of these cell-free DNA fragments originates from specific chromosomes, especially chromosomes 1 and 9. In healthy adult somatic cells, the centromeric and pericentromeric regions of these chromosomes are normally densely methylated. However, in many cancer types, these regions are preferentially hypomethylated. This can lead to double-stranded DNA breaks or it can directly impair the formation of proper kinetochore structures. This type of chromosomal instability is a precursor to the formation of nuclear anomalies, including lagging chromosomes and anaphase bridges. DNA fragments derived from these structures can recruit their own nuclear envelope and form secondary nuclear structures known as micronuclei, which can localize to the nuclear periphery and bud out from the membrane. We postulate that the majority of cell-free DNA present in the growth medium of cultured 143B cells originates from these micronuclei. |
| format | Article |
| id | doaj-art-97cf857cfc46473ca7562642d2ea3b1a |
| institution | Kabale University |
| issn | 1423-0380 |
| language | English |
| publishDate | 2018-09-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Tumor Biology |
| spelling | doaj-art-97cf857cfc46473ca7562642d2ea3b1a2025-08-20T03:56:55ZengSAGE PublishingTumor Biology1423-03802018-09-014010.1177/1010428318801190Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cellsAbel Jacobus Bronkhorst0Johannes F Wentzel1Vida Ungerer2Dimetrie L Peters3Janine Aucamp4Etienne Pierre de Villiers5Stefan Holdenrieder6Piet J Pretorius7Institute for Laboratory Medicine, German Heart Centre, Technical University Munich, Munich, GermanyCentre of Excellence for Nutrition (CEN), North-West University, Potchefstroom, South AfricaInstitute for Laboratory Medicine, German Heart Centre, Technical University Munich, Munich, GermanyHuman Metabolomics, Biochemistry Division, North-West University, Potchefstroom, South AfricaCentre of Excellence for Pharmaceutical Sciences, North-West University, Potchefstroom, South AfricaCentre for Tropical Medicine, University of Oxford, Oxford, United KingdomInstitute for Laboratory Medicine, German Heart Centre, Technical University Munich, Munich, GermanyHuman Metabolomics, Biochemistry Division, North-West University, Potchefstroom, South AfricaThe true importance of cell-free DNA in human biology, together with the potential scale of its clinical utility, is tarnished by a lack of understanding of its composition and origin. In investigating the cell-free DNA present in the growth medium of cultured 143B cells, we previously demonstrated that the majority of cell-free DNA is neither a product of apoptosis nor necrosis. In the present study, we investigated the composition and origin of this cell-free DNA population using next-generation sequencing. We found that the cell-free DNA comprises mainly of repetitive DNA, including α-satellite DNA, mini satellites, and transposons that are currently active or exhibit the capacity to become reactivated. A significant portion of these cell-free DNA fragments originates from specific chromosomes, especially chromosomes 1 and 9. In healthy adult somatic cells, the centromeric and pericentromeric regions of these chromosomes are normally densely methylated. However, in many cancer types, these regions are preferentially hypomethylated. This can lead to double-stranded DNA breaks or it can directly impair the formation of proper kinetochore structures. This type of chromosomal instability is a precursor to the formation of nuclear anomalies, including lagging chromosomes and anaphase bridges. DNA fragments derived from these structures can recruit their own nuclear envelope and form secondary nuclear structures known as micronuclei, which can localize to the nuclear periphery and bud out from the membrane. We postulate that the majority of cell-free DNA present in the growth medium of cultured 143B cells originates from these micronuclei.https://doi.org/10.1177/1010428318801190 |
| spellingShingle | Abel Jacobus Bronkhorst Johannes F Wentzel Vida Ungerer Dimetrie L Peters Janine Aucamp Etienne Pierre de Villiers Stefan Holdenrieder Piet J Pretorius Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells Tumor Biology |
| title | Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells |
| title_full | Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells |
| title_fullStr | Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells |
| title_full_unstemmed | Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells |
| title_short | Sequence analysis of cell-free DNA derived from cultured human bone osteosarcoma (143B) cells |
| title_sort | sequence analysis of cell free dna derived from cultured human bone osteosarcoma 143b cells |
| url | https://doi.org/10.1177/1010428318801190 |
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