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Published in: Cancer Cell International 1/2006

Open Access 01-12-2006 | Primary research

Qualitative and quantitative characteristics of the extracellular DNA delivered to the nucleus of a living cell

Authors: Vladimir A Rogachev, Anastasia Likhacheva, Oksana Vratskikh, Lyudmila V Mechetina, Tamara E Sebeleva, Sergei S Bogachev, Leonid A Yakubov, Mikhail A Shurdov

Published in: Cancer Cell International | Issue 1/2006

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Abstract

Background

The blood plasma and other intertissue fluids usually contain a certain amount of DNA, getting there due to a natural cell death in the organism. Cells of this organism can capture the extracellular DNA, whereupon it is delivered to various cell compartments. It is hypothesized that the extracellular DNA is involved in the transfer of genetic information and its fixation in the genome of recipient cell.

Results

The existence of an active flow of extracellular DNA into the cell is demonstrated using human breast adenocarcinoma (MCF-7) cells as a recipient culture. The qualitative state of the DNA fragments delivered to the main cell compartments (cytoplasm and interchromosomal fraction) was assessed. The extracellular DNA delivered to the cell is characterized quantitatively.

Conclusion

It is demonstrated that the extracellular DNA fragments in several minutes reach the nuclear space, where they are processed so that their linear size increases from about 500 bp to 10,000 bp. The amount of free extracellular DNA fragments simultaneously present in the nuclear space may reach up to 2% of the haploid genome. Using individual DNA fragments with a known molecular weight and sequence as an extracellular DNA, it is found that these fragments degrade instantly in the culture liquid in the absence of a competitor DNA and are delivered into the cell as degradants. When adding a sufficient amount of competitor DNA, the initial undegraded molecules of the DNA fragments with the known molecular weight and sequence are detectable both in the cytoplasm and nuclear space only at the zero point of experiments. The labeled precursor α-dNTP*, added to culture medium, was undetectable inside the cell in all the experiments.
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Literature
1.
go back to reference Anker P: Quantitative aspects of plasma/serum DNA in cancer patients. Ann New York Aca Sci. 2000, 906: 5-7.CrossRef Anker P: Quantitative aspects of plasma/serum DNA in cancer patients. Ann New York Aca Sci. 2000, 906: 5-7.CrossRef
2.
go back to reference Anker P, Mulcahy H, Chen XQ, Stroun M: Detection of circulating tumour DNA in the blood (plasma/serum) of cancer patients. Cancer Metast Rev. 1999, 18: 65-73. 10.1023/A:1006260319913.CrossRef Anker P, Mulcahy H, Chen XQ, Stroun M: Detection of circulating tumour DNA in the blood (plasma/serum) of cancer patients. Cancer Metast Rev. 1999, 18: 65-73. 10.1023/A:1006260319913.CrossRef
3.
go back to reference Anker P, Mulcahy H, Stroun M: Circulating nucleic acids in plasma and serum as a noninvasive investigation for cancer: Time for large-scale clinical studies?. Int J Cancer. 2003, 103: 149-152. 10.1002/ijc.10791.CrossRefPubMed Anker P, Mulcahy H, Stroun M: Circulating nucleic acids in plasma and serum as a noninvasive investigation for cancer: Time for large-scale clinical studies?. Int J Cancer. 2003, 103: 149-152. 10.1002/ijc.10791.CrossRefPubMed
4.
go back to reference Ledoux L: Uptake of DNA by living cells. Prog Nucl Acid Res Mol Biol. 1965, 4: 231-267.CrossRef Ledoux L: Uptake of DNA by living cells. Prog Nucl Acid Res Mol Biol. 1965, 4: 231-267.CrossRef
5.
go back to reference Anker P, Jachertz D, Stroun M, Brögger R, Lederrey C, Henri J, Maurice PA: The role of extracellular DNA in the transfer of information from T to B human lymphocytes in the course of an immune response. J Immunogenet. 1980, 7: 475-481.CrossRefPubMed Anker P, Jachertz D, Stroun M, Brögger R, Lederrey C, Henri J, Maurice PA: The role of extracellular DNA in the transfer of information from T to B human lymphocytes in the course of an immune response. J Immunogenet. 1980, 7: 475-481.CrossRefPubMed
6.
go back to reference Bergsmedh A, Szeles A, Spetz AL, Holmgren L: Loss of the p21(Cip1/Waf1) cyclin kinase inhibitor results in propagation of horizontally transferred DNA. Cancer Res. 2002, 62: 575-579.PubMed Bergsmedh A, Szeles A, Spetz AL, Holmgren L: Loss of the p21(Cip1/Waf1) cyclin kinase inhibitor results in propagation of horizontally transferred DNA. Cancer Res. 2002, 62: 575-579.PubMed
7.
go back to reference Holmgren L, Szeles A, Rajnavolgyi E, Folkman J, Klein G, Ernberg I, Falk LI: Horizontal transfer of DNA by the uptake of apoptotic bodies. Blood. 1999, 93: 3956-3963.PubMed Holmgren L, Szeles A, Rajnavolgyi E, Folkman J, Klein G, Ernberg I, Falk LI: Horizontal transfer of DNA by the uptake of apoptotic bodies. Blood. 1999, 93: 3956-3963.PubMed
8.
go back to reference Garcia-Olmo D, Ontanon J, Garcia-Olmo DC, Atienzar M, Valleo M: Detection of genomically-tagged cancer cells in different tissues at different stages of tumor development: lack of correlation with the formation of metastasis. Cancer Lett. 1999, 140: 11-20. 10.1016/S0304-3835(99)00046-4.CrossRefPubMed Garcia-Olmo D, Ontanon J, Garcia-Olmo DC, Atienzar M, Valleo M: Detection of genomically-tagged cancer cells in different tissues at different stages of tumor development: lack of correlation with the formation of metastasis. Cancer Lett. 1999, 140: 11-20. 10.1016/S0304-3835(99)00046-4.CrossRefPubMed
9.
go back to reference Garcia-Olmo D, Garcia-Olmo DC, Ontanon J, Martinez E: Horizontal transfer of DNA and the "genometastasis hypothesis". Blood. 2000, 95: 724-725.PubMed Garcia-Olmo D, Garcia-Olmo DC, Ontanon J, Martinez E: Horizontal transfer of DNA and the "genometastasis hypothesis". Blood. 2000, 95: 724-725.PubMed
10.
go back to reference Yakubov LA, Petrova NA, Popova NA, Semenov DV, Nikolin VP, Os'kina IN: The role of extracellular DNA in the stability and variability of cell genomes. Doklady Biochemistry and Biophysics. 2002, 382: 406-410. 10.1023/A:1014403223031.CrossRef Yakubov LA, Petrova NA, Popova NA, Semenov DV, Nikolin VP, Os'kina IN: The role of extracellular DNA in the stability and variability of cell genomes. Doklady Biochemistry and Biophysics. 2002, 382: 406-410. 10.1023/A:1014403223031.CrossRef
12.
go back to reference Loke SL, Stein CA, Zhang XH, Moir K, Nakanishi M, Subasinghe C, Cohen JS, Neckers LM: Characterization of oligonucleotide transport into living cells. Proc Natl Acad Sci USA. 1989, 86: 3474-3478. 10.1073/pnas.86.10.3474.PubMedCentralCrossRefPubMed Loke SL, Stein CA, Zhang XH, Moir K, Nakanishi M, Subasinghe C, Cohen JS, Neckers LM: Characterization of oligonucleotide transport into living cells. Proc Natl Acad Sci USA. 1989, 86: 3474-3478. 10.1073/pnas.86.10.3474.PubMedCentralCrossRefPubMed
13.
go back to reference Zamecnik P, Aghajanian J, Zamecnik M, Goodchild J, Witman G: Electron micrographic studies of transport of oligodeoxynucleotides across eukaryotic membranes. Proc Natl Acad Sci USA. 1994, 91: 3156-3160. 10.1073/pnas.91.8.3156.PubMedCentralCrossRefPubMed Zamecnik P, Aghajanian J, Zamecnik M, Goodchild J, Witman G: Electron micrographic studies of transport of oligodeoxynucleotides across eukaryotic membranes. Proc Natl Acad Sci USA. 1994, 91: 3156-3160. 10.1073/pnas.91.8.3156.PubMedCentralCrossRefPubMed
14.
go back to reference Shestova OE, Andreeva AYu, Vlasov VV, Yakubov LA: Transport of complexes of oligonucleotides with cell-surface proteins to the cell nucleus. Dokl Ross Akad Nauk. 1999, 368: 264-267. Shestova OE, Andreeva AYu, Vlasov VV, Yakubov LA: Transport of complexes of oligonucleotides with cell-surface proteins to the cell nucleus. Dokl Ross Akad Nauk. 1999, 368: 264-267.
15.
go back to reference Ledoux L, Charles P: Fate of exogenous DNA in mammals. Uptake of Informative Molecules by Living Cells. Edited by: Ledoux L. 1972, North-Holland Publishing Company, Amsterdam, 397-413. Ledoux L, Charles P: Fate of exogenous DNA in mammals. Uptake of Informative Molecules by Living Cells. Edited by: Ledoux L. 1972, North-Holland Publishing Company, Amsterdam, 397-413.
16.
go back to reference Yakubov LA, Deeva EA, Zarytova VF, Ivanova EM, Ryte AS, Yurchenko LY, Vlassov VV: Mechanisms of oligonucleotide uptake by cells: Involvement of specific receptors?. Proc Natl Acad Sci USA. 1989, 86: 6454-6458. 10.1073/pnas.86.17.6454.PubMedCentralCrossRefPubMed Yakubov LA, Deeva EA, Zarytova VF, Ivanova EM, Ryte AS, Yurchenko LY, Vlassov VV: Mechanisms of oligonucleotide uptake by cells: Involvement of specific receptors?. Proc Natl Acad Sci USA. 1989, 86: 6454-6458. 10.1073/pnas.86.17.6454.PubMedCentralCrossRefPubMed
17.
go back to reference Koulintchenko M, Konstantinov Y, Dietrich A: Plant mitochondria activity import DNA via the permeability transition pore complex. EMBO J. 2003, 22: 1245-1254. 10.1093/emboj/cdg128.PubMedCentralCrossRefPubMed Koulintchenko M, Konstantinov Y, Dietrich A: Plant mitochondria activity import DNA via the permeability transition pore complex. EMBO J. 2003, 22: 1245-1254. 10.1093/emboj/cdg128.PubMedCentralCrossRefPubMed
18.
go back to reference Baricheva EM, Berrios M, Bogachev SS, Borisevich IV, Lapik ER, Sharakhov IV, Stuurmman N, Fisher PA: DNA from Drosophila melanogaster β-heterochromatin binds specifically to nuclear lamina in vitro and the nuclear envelope in situ. Gene. 1996, 171: 171-176. 10.1016/0378-1119(96)00002-9.CrossRefPubMed Baricheva EM, Berrios M, Bogachev SS, Borisevich IV, Lapik ER, Sharakhov IV, Stuurmman N, Fisher PA: DNA from Drosophila melanogaster β-heterochromatin binds specifically to nuclear lamina in vitro and the nuclear envelope in situ. Gene. 1996, 171: 171-176. 10.1016/0378-1119(96)00002-9.CrossRefPubMed
19.
go back to reference Lees-Miller SP, Meek K: Repair of DNA double strand breaks by non-homologous end joining. Biochimie. 2003, 85: 1161-1173. 10.1016/j.biochi.2003.10.011.CrossRefPubMed Lees-Miller SP, Meek K: Repair of DNA double strand breaks by non-homologous end joining. Biochimie. 2003, 85: 1161-1173. 10.1016/j.biochi.2003.10.011.CrossRefPubMed
20.
go back to reference Likhacheva A, Vratskikh O, Rogachev VA, Mechetina LV, Sebeleva TE, Bogachev SS, Yakubov LA, Shurdov MA: Natural human gene correction by small extracellular genomic DNA fragments. Proceedings of EMBO workshop "The functional organization of the cell nucleus: 5–8 May 2006 Prague. 2006, 68: 33- Likhacheva A, Vratskikh O, Rogachev VA, Mechetina LV, Sebeleva TE, Bogachev SS, Yakubov LA, Shurdov MA: Natural human gene correction by small extracellular genomic DNA fragments. Proceedings of EMBO workshop "The functional organization of the cell nucleus: 5–8 May 2006 Prague. 2006, 68: 33-
21.
go back to reference Glover D: DNA Cloning: A Practical Approach. 1985, IRL Press, Oxford Glover D: DNA Cloning: A Practical Approach. 1985, IRL Press, Oxford
22.
go back to reference Roberts DB: Drosophila: A Practical Approach. 1986, IRL Press, Oxford Roberts DB: Drosophila: A Practical Approach. 1986, IRL Press, Oxford
Metadata
Title
Qualitative and quantitative characteristics of the extracellular DNA delivered to the nucleus of a living cell
Authors
Vladimir A Rogachev
Anastasia Likhacheva
Oksana Vratskikh
Lyudmila V Mechetina
Tamara E Sebeleva
Sergei S Bogachev
Leonid A Yakubov
Mikhail A Shurdov
Publication date
01-12-2006
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2006
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/1475-2867-6-23

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