CoChat
Sign Up
Cited by 7Carcinogens and Genotoxicity AssessmentImmunotherapy and Immune ResponsesImmunotoxicology and immune responses

A Mouse Strain Defective in Both T Cells and NK Cells Has Enhanced Sensitivity to Tumor Induction by Plasmid DNA Expressing Both Activated H-Ras and c-Myc

Open access
Sheng-Fowler, Li, Tu, Wei, Fu, Haiqing, Murata, Haruhiko, Lanning, Lynda, Foseh, Gideon, Macauley, Juliete, Blair, Donald +4 more·2014·PLoS ONE
DOI 10.1371/journal.pone.0108926PMC PMC4193875
Open this paper in CoChat — free accountOpen PDF

Abstract

As part of safety studies to evaluate the risk of residual cellular DNA in vaccines manufactured in tumorigenic cells, we have been developing in vivo assays to detect and quantify the oncogenic activity of DNA. We generated a plasmid expressing both an activated human H-ras gene and murine c-myc gene and showed that 1 µg of this plasmid, pMSV-T24-H-ras/MSV-c-myc, was capable of inducing tumors in newborn NIH Swiss mice. However, to be able to detect the oncogenicity of dominant activated oncogenes in cellular DNA, a more sensitive system was needed. In this paper, we demonstrate that the newborn CD3 epsilon transgenic mouse, which is defective in both T-cell and NK-cell functions, can detect the oncogenic activity of 25 ng of the circular form of pMSV-T24-H-ras/MSV-c-myc. When this plasmid was inoculated as linear DNA, amounts of DNA as low as 800 pg were capable of inducing tumors. Animals were found that had multiple tumors, and these tumors were independent and likely clonal. These results demonstrate that the newborn CD3 epsilon mouse is highly sensitive for the detection of oncogenic activity of DNA. To determine whether it can detect the oncogenic activity of cellular DNA derived from four human tumor-cell lines (HeLa, A549, HT-1080, and CEM), DNA (100 µg) was inoculated into newborn CD3 epsilon mice both in the presence of 1 µg of linear pMSV-T24-H-ras/MSV-c-myc as positive control and in its absence. While tumors were induced in 100% of mice with the positive-control plasmid, no tumors were induced in mice receiving any of the tumor DNAs alone. These results demonstrate that detection of oncogenes in cellular DNA derived from four human tumor-derived cell lines in this mouse system was not possible; the results also show the importance of including a positive-control plasmid to detect inhibitory effects of the cellular DNA.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0108926

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 48

  1. [1]Petricciani JC, Horaud FN (1995) DNA, dragons and sanity. Biologicals 23: 233– 238. 8527124
  2. [2]Petricciani J, Loewer J (2001) An overview of cell DNA issues. Dev Biol 106: 275– 282. 11761240
  3. [3]Lewis AM Jr, Krause P, Peden K (2001) A defined-risks approach to the regulatory assessment of the use of neoplastic cells as substrates for viral vaccine manufacture. Dev Biol 106: 513– 535. 11761266
  4. [4]Sheng-Fowler L, Lewis AM Jr, Peden K (2009) Issues associated with residual cell-substrate DNA in viral vaccines. Biologicals 37: 190– 195. 19285882
  5. [5]Sheng L, Cai F, Zhu Y, Pal A, Athanasiou M, et al (2008) Oncogenicity of DNA in vivo: Tumor induction with expression plasmids for activated H-ras and c-myc. Biologicals 36: 184– 197. 18218323
  6. [6]Land H, Parada LF, Weinberg RA (1983) Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304: 596– 602. 6308472
  7. [7]Santos E, Pulciani S, Barbacid M (1984) Characterization of a human transforming gene isolated from T24 bladder carcinoma cells. Fed Proc 43: 2280– 2286. 6325252
  8. [8]Thompson TC, Southgate J, Kitchener G, Land H (1989) Multistage carcinogenesis induced by ras and myc oncogenes in a reconstituted organ. Cell 56: 917– 930. 2538247
  9. [9]Small MB, Hay N, Schwab M, Bishop JM (1987) Neoplastic transformation by the human gene N-myc. Mol Cell Biol 7: 1638– 1645. 3299052 PMC365263
  10. [10]Zerlin M, Julius MA, Cerni C, Marcu KB (1987) Elevated expression of an exogenous c-myc gene is insufficient for transformation and tumorigenic conversion of established fibroblasts. Oncogene 1: 19– 27. 3325875
  11. [11]Sheng-Fowler L, Cai F, Fu H, Zhu Y, Orrison B, et al (2010) Tumors induced in mice by direct inoculation of plasmid DNA expressing both activated H-ras and c-myc. Int J Biol Sci 6: 151– 162. 20376206 PMC2850538
  12. [12]Wang B, Biron C, She J, Higgins K, Sunshine MJ, et al (1994) A block in both early T lymphocyte and natural killer cell development in transgenic mice with high-copy numbers of the human CD3E gene. Proc Natl Acad Sci U S A 91: 9402– 9406. 7937778 PMC44820
  13. [14]Gey GO (1954) Some aspects of the constitution and behavior of normal and malignant cells maintained in continuous culture. Harvey Lect 50: 154– 229. 13306037
  14. [15]Gey GO, Bang FB, Gey MK (1954) Responses of a variety of normal and malignant cells to continuous cultivation, and some practical applications of these responses to problems in the biology of disease. Ann N Y Acad Sci 58: 976– 999. 14350495
  15. [16]Lieber M, Smith B, Szakal A, Nelson-Rees W, Todaro G (1976) A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int J Cancer 17: 62– 70. 175022
  16. [17]Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, et al (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51: 1417– 1423. 4357758
  17. [18]Rasheed S, Nelson-Rees WA, Toth EM, Arnstein P, Gardner MB (1974) Characterization of a newly derived human sarcoma cell line (HT-1080). Cancer 33: 1027– 1033. 4132053
  18. [19]Foley GE, Lazarus H, Farber S, Uzman BG, Boone BA, et al (1965) Continuous culture of human lymphoblasts from peripheral blood of a child with acute leukemia. Cancer 18: 522– 529. 14278051
  19. [20]Routes JM, Ryan S, Li H, Steinke J, Cook JL (2000) Dissimilar immunogenicities of human papillomavirus E7 and adenovirus E1A proteins influence primary tumor development. Virology 277: 48– 57. 11062035
  20. [21]Israel MA, Simmons DT, Hourihan SL, Rowe WP, Martin MA (1979) Interrupting the early region of polyoma virus DNA enhances tumorigenicity. Proc Natl Acad Sci U S A 76: 3713– 3716. 226975 PMC383903
  21. [22]Bouchard L, Gelinas C, Asselin C, Bastin M (1984) Tumorigenic activity of polyoma virus and SV40 DNAs in newborn rodents. Virology 135: 53– 64. 6328753
  22. [23]Sheng-Fowler L, Lewis AM Jr, Peden K (2009) Quantitative determination of the infectivity of the proviral DNA of a retrovirus in vitro: Evaluation of methods for DNA inactivation. Biologicals 37: 259– 269. 19457682
  23. [24]Cupillard L, Juillard V, Latour S, Colombet G, Cachet N, et al (2005) Impact of plasmid supercoiling on the efficacy of a rabies DNA vaccine to protect cats. Vaccine 23: 1910– 1916. 15734063
  24. [25]Pillai VB, Hellerstein M, Yu T, Amara RR, Robinson HL (2008) Comparative studies on in vitro expression and in vivo immunogenicity of supercoiled and open circular forms of plasmid DNA vaccines. Vaccine 26: 1136– 1141. 18242791 PMC2692637
  25. [26]Wolff JA, Ludtke JJ, Acsadi G, Williams P, Jani A (1992) Long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum Mol Genet 1: 363– 369. 1301910
  26. [27]Zietman AL, Suit HD, Ramsay JR, Silobrcic V, Sedlacek RS (1988) Quantitative studies on the transplantability of murine and human tumors into the brain and subcutaneous tissues of NCr/Sed nude mice. Cancer Res 48: 6510– 6516. 3052803
  27. [28]Hill RP (1980) An appraisal of in vivo assays of excised tumours. Br J Cancer Suppl 4: 230–239. PMC2149236 6932930
  28. [29]Lewis AM, Alling DW, Banks SM, Soddu S, Cook JL (1999) Evaluating virus-transformed cell tumorigenicity. J Virol Methods 79: 41– 50. 10328534
  29. [30]Boshart M, Gissmann L, Ikenberg H, Kleinheinz A, Scheurlen W, et al (1984) A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. Embo j 3: 1151– 1157. 6329740 PMC557488
  30. [31]Foley GE, Handler AH, Lynch PM, Wolman SR, Stulberg CS, et al (1965) Loss of neoplastic properties in vitro. II. Observations on KB sublines. Cancer Res 25: 1254– 1261. 5839717
  31. [32]Merl S, Kloster B, Moore J, Hubbell C, Tomar R, et al (1984) Efficient transformation of previously activated and dividing T lymphocytes by human T cell leukemia-lymphoma virus. Blood 64: 967– 974. 6333258
  32. [33]Mortimer I, Tam P, MacLachlan I, Graham RW, Saravolac EG, et al (1999) Cationic lipid-mediated transfection of cells in culture requires mitotic activity. Gene Ther 6: 403– 411. 10435090
  33. [34]Zabner J, Fasbender AJ, Moninger T, Poellinger KA, Welsh MJ (1995) Cellular and molecular barriers to gene transfer by a cationic lipid. J Biol Chem 270: 18997– 19007. 7642560
  34. [35]Peden KWC, Pipas JM, Pearson-White S, Nathans D (1980) Isolation of mutants of an animal virus in bacteria. Science 209: 1392– 1396. 6251547
  35. [36]Israel MA, Chan HW, Hourihan SL, Rowe WP, Martin MA (1979) Biological activity of polyoma viral DNA in mice and hamsters. J Virol 29: 990– 996. 221686 PMC353259
  36. [37]Hesketh R (1997) The Oncogene and Tumour Suppressor Gene FactsBook. London: Academic Press. 549 p.
  37. [38]Liu C, Yu J, Yu S, Lavker RM, Cai L, et al (2010) MicroRNA-21 acts as an oncomir through multiple targets in human hepatocellular carcinoma. J Hepatol 53: 98– 107. 20447717
  38. [39]Cowland JB, Hother C, Gronbaek K (2007) MicroRNAs and cancer. Apmis 115: 1090– 1106. 18042145
  39. [40]Iorio MV, Croce CM (2012) MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive review. EMBO Mol Med 4: 143– 159. 22351564 PMC3376845
  40. [41]Kong YW, Ferland-McCollough D, Jackson TJ, Bushell M (2012) microRNAs in cancer management. Lancet Oncol 13: e249– 258. 22652233
  41. [42]Macville M, Schrock E, Padilla-Nash H, Keck C, Ghadimi BM, et al (1999) Comprehensive and definitive molecular cytogenetic characterization of HeLa cells by spectral karyotyping. Cancer Res 59: 141– 150. 9892199
  42. [43]Meissner JD (1999) Nucleotide sequences and further characterization of human papillomavirus DNA present in the CaSki, SiHa and HeLa cervical carcinoma cell lines. J Gen Virol 80 (Pt 7): 1725– 1733. 10423141
  43. [44]Chang H, Jackson DG, Kayne PS, Ross-Macdonald PB, Ryseck RP, et al (2011) Exome sequencing reveals comprehensive genomic alterations across eight cancer cell lines. PLoS ONE 6: e21097. 21701589 PMC3118809
  44. [45]Marshall CJ, Hall A, Weiss RA (1982) A transforming gene present in human sarcoma cell lines. Nature 299: 171– 173. 6287287
  45. [46]Hall A, Marshall CJ, Spurr NK, Weiss RA (1983) Identification of transforming gene in two human sarcoma cell lines as a new member of the ras gene family located on chromosome 1. Nature 303: 396– 400. 6304521
  46. [47]Brown R, Marshall CJ, Pennie SG, Hall A (1984) Mechanism of activation of an N-ras gene in the human fibrosarcoma cell line HT1080. Embo J 3: 1321– 1326. 6086315 PMC557516
  47. [48]Pulciani S, Santos E, Lauver AV, Long LK, Robbins KC, et al (1982) Oncogenes in human tumor cell lines: molecular cloning of a transforming gene from human bladder carcinoma cells. Proc Natl Acad Sci U S A 79: 2845– 2849. 6953433 PMC346303
  48. [49]Blair DG, Cooper CS, Oskarsson MK, Eader LA, Vande Woude GF (1982) New method for detecting cellular transforming genes. Science 218: 1122– 1125. 6293052

Cited by

Sorted by citations

Authors
Sheng-Fowler, L., Tu, W., Fu, H., Murata, H., Lanning, L., Foseh, G., Macauley, J., Blair, D., Hughes, S. H., Coffin, J. M., Lewis, A. M., & Peden, K.
Affiliations
  • Center for Biologics Evaluation and Research (US)
  • National Institute of Allergy and Infectious Diseases (US)
  • National Cancer Institute (US)
  • Frederick National Laboratory for Cancer Research (US)
  • Tufts University (US)
Corresponding author
Peden, Keith
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-10-10
Volume
9
Issue
10
Pages
e108926
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0108926
PMC
PMC4193875
OpenAlex
W2064155411
Cited by
7
References
0
Citations by year
2025: 1 · 2024: 2 · 2023: 1 · 2020: 2 · 2019: 1
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, public-domain)
  • PubMed Central (publishedVersion, public-domain)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Department of Health and Human Services (HHSN26120080001E)
  • National Institutes of Health (HHSN26120080001E)
  • U.S. Public Health Service (HHSN 26120080001E)
  • National Cancer Institute (HHSN26120080001E)
  • National Institute of Allergy and Infectious Diseases (Y01 AI004893, YI-AI-4893-02NIAID)
  • Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
Keywords
T cell deficiency, NK cell deficiency, H-Ras, c-Myc, plasmid DNA, tumor induction, vaccine safety, linear DNA, human cancer cell lines, HeLa, A549 cells, CEM
MeSH terms
Animals, DNA, Circular, DNA, Neoplasm, Humans, Killer Cells, Natural, Mice, Transgenic, Neoplasms, Plasmids, Genes, ras, T-Lymphocytes, Genes, myc, Cell Line, Tumor, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review