Contributions of viral oncogenes of HPV-18 and hypoxia to oxidative stress and genetic damage in human keratinocytes.

Female Humans Human papillomavirus 18 / genetics Reactive Oxygen Species / metabolism Oncogene Proteins, Viral / genetics Oxidative Stress / genetics Keratinocytes / metabolism Oncogenes Hypoxia / metabolism Papillomavirus E7 Proteins / genetics Uterine Cervical Neoplasms / pathology Papillomavirus Infections / genetics Tumor Microenvironment

Journal

Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288

Informations de publication

Date de publication:
18 10 2023
Historique:
received: 17 08 2023
accepted: 12 10 2023
medline: 23 10 2023
pubmed: 19 10 2023
entrez: 18 10 2023
Statut: epublish

Résumé

Infection with high-risk human papillomaviruses like HPV-16 and HPV-18 is highly associated with the development of cervical and other cancers. Malignant transformation requires viral oncoproteins E5, E6 and E7, which promote cell proliferation and increase DNA damage. Oxidative stress and hypoxia are also key factors in cervical malignant transformation. Increased levels of reactive species of oxygen (ROS) and nitrogen (RNS) are found in the hypoxic tumor microenvironment, promoting genetic instability and invasiveness. In this work, we studied the combined effect of E5, E6 and E7 and hypoxia in increasing oxidative stress and promoting DNA damage and nuclear architecture alterations. HaCaT cells containing HPV-18 viral oncogenes (HaCaT E5/E6/E7-18) showed higher ROS levels in normoxia and higher levels of RNS in hypoxia compared to HaCaT parental cells, as well as higher genetic damage in hypoxia as measured by γH2AX and comet assays. In hypoxia, HaCaT E5/E6/E7-18 increased its nuclear dry mass and both cell types displayed marked heterogeneity in nuclear dry mass distribution and increased nuclear foci. Our results show contributions of both viral oncogenes and hypoxia to oxidative stress, DNA damage and altered nuclear architecture, exemplifying how an altered microenvironment combines with oncogenic transformation to promote tumor progression.

Identifiants

DOI: 10.1038/s41598-023-44880-3 PMID: 37853061 PMC: PMC10584980
pubmed: 37853061
doi: 10.1038/s41598-023-44880-3
pii: 10.1038/s41598-023-44880-3
pmc: PMC10584980
doi:

Substances chimiques

Reactive Oxygen Species 0
Oncogene Proteins, Viral 0
Papillomavirus E7 Proteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

17734

Informations de copyright

© 2023. Springer Nature Limited.

Références

Pal, A. & Kundu, R. Human papillomavirus E6 and E7: The cervical cancer hallmarks and targets for therapy. Front. Microbiol. 21(10), 3116. https://doi.org/10.3389/fmicb.2019.03116 (2020).
doi: 10.3389/fmicb.2019.03116
de Sanjose, S. et al. Retrospective international survey and HPV time trends study group. Human papillomavirus genotype attribution in invasive cervical cancer: A retrospective cross-sectional worldwide study. Lancet Oncol. 11(11), 1048–56. https://doi.org/10.1016/S1470-2045(10)70230-8 (2010).
doi: 10.1016/S1470-2045(10)70230-8 pubmed: 20952254
Venuti, A. et al. Papillomavirus E5: The smallest oncoprotein with many functions. Mol. Cancer. 11(10), 140. https://doi.org/10.1186/1476-4598-10-140 (2011).
doi: 10.1186/1476-4598-10-140
Bouvard, V., Matlashewski, G., Gu, Z. M., Storey, A. & Banks, L. The human papillomavirus type 16 E5 gene cooperates with the E7 gene to stimulate proliferation of primary cells and increases viral gene expression. Virology 203(1), 73–80. https://doi.org/10.1006/viro.1994.1456 (1994).
doi: 10.1006/viro.1994.1456 pubmed: 8030286
Lechner, M. S. et al. Human papillomavirus E6 proteins bind p53 in vivo and abrogate p53-mediated repression of transcription. EMBO J. 11(8), 3045–3052. https://doi.org/10.1002/j.1460-2075.1992.tb05375.x. (1992).
doi: 10.1002/j.1460-2075.1992.tb05375.x. pubmed: 1379175 pmcid: 556787
Scheffner, M., Huibregtse, J. M., Vierstra, R. D. & Howley, P. M. The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53. Cell 75(3), 495–505. https://doi.org/10.1016/0092-8674(93)90384-3 (1993).
doi: 10.1016/0092-8674(93)90384-3 pubmed: 8221889
Dyson, N., Howley, P. M., Münger, K. & Harlow, E. The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product. Science 243(4893), 934–937. https://doi.org/10.1126/science.2537532 (1989).
doi: 10.1126/science.2537532 pubmed: 2537532
Hussain, S. P. & Harris, C. C. Inflammation and cancer: An ancient link with novel potentials. Int. J. Cancer 121(11), 2373–2380. https://doi.org/10.1002/ijc.23173 (2007).
doi: 10.1002/ijc.23173 pubmed: 17893866
Kawanishi, S., Ohnishi, S., Ma, N., Hiraku, Y. & Murata, M. Crosstalk between DNA Damage and Inflammation in the multiple steps of carcinogenesis. Int. J. Mol. Sci. 18(8), 1808. https://doi.org/10.3390/ijms18081808 (2017).
doi: 10.3390/ijms18081808 pubmed: 28825631 pmcid: 5578195
Prati, B., Marangoni, B. & Boccardo, E. Human papillomavirus and genome instability: From productive infection to cancer. Clinics (Sao Paulo) 73(suppl 1), e539s. https://doi.org/10.6061/clinics/2018/e539s (2018).
doi: 10.6061/clinics/2018/e539s pubmed: 30208168
Cruz-Gregorio, A. et al. Human papillomavirus types 16 and 18 early-expressed proteins differentially modulate the cellular redox state and DNA damage. Int. J. Biol. Sci. 14(1), 21–35. https://doi.org/10.7150/ijbs.21547 (2018).
doi: 10.7150/ijbs.21547 pubmed: 29483822 pmcid: 5821046
Pasha, A. et al. Inhibition of inducible nitric oxide synthase (iNOS) by andrographolide and in vitro evaluation of its antiproliferative and proapoptotic effects on cervical cancer. Oxid. Med. Cell Longev. 2021, 6692628. https://doi.org/10.1155/2021/6692628 (2021).
doi: 10.1155/2021/6692628 pubmed: 33815659 pmcid: 8010528
Rahkola, P., Mikkola, T. S., Ylikorkala, O. & Vaisanen-Tommiska, M. Association between high risk papillomavirus DNA and nitric oxide release in the human uterine cervix. Gynecol. Oncol. 114(2), 323–326. https://doi.org/10.1016/j.ygyno.2009.05.003 (2009).
doi: 10.1016/j.ygyno.2009.05.003 pubmed: 19481240
Hernansanz-Agustín, P. et al. Acute hypoxia produces a superoxide burst in cells. Free Radic. Biol. Med. 71, 146–156. https://doi.org/10.1016/j.freeradbiomed.2014.03.011 (2014).
doi: 10.1016/j.freeradbiomed.2014.03.011 pubmed: 24637263
Guzy, R. D. & Schumacker, P. T. Oxygen sensing by mitochondria at complex III: The paradox of increased reactive oxygen species during hypoxia. Exp. Physiol. 91(5), 807–819. https://doi.org/10.1113/expphysiol.2006.033506 (2006).
doi: 10.1113/expphysiol.2006.033506 pubmed: 16857720
Gillies, R. J., Verduzco, D. & Gatenby, R. A. Evolutionary dynamics of carcinogenesis and why targeted therapy does not work. Nat. Rev. Cancer 12(7), 487–493. https://doi.org/10.1038/nrc3298 (2012).
doi: 10.1038/nrc3298 pubmed: 22695393 pmcid: 4122506
Shimojo, Y. et al. Attenuation of reactive oxygen species by antioxidants suppresses hypoxia-induced epithelial-mesenchymal transition and metastasis of pancreatic cancer cells. Clin. Exp. Metastasis 30(2), 143–154. https://doi.org/10.1007/s10585-012-9519-8 (2013).
doi: 10.1007/s10585-012-9519-8 pubmed: 22833345
Kawanishi, S. et al. Nitrative and oxidative DNA damage in infection-related carcinogenesis in relation to cancer stem cells. Genes Environ. 1(38), 26. https://doi.org/10.1186/s41021-016-0055-7 (2017).
doi: 10.1186/s41021-016-0055-7
Kawanishi, S. & Hiraku, Y. Oxidative and nitrative DNA damage as biomarker for carcinogenesis with special reference to inflammation. Antioxid. Redox. Signal. 8(5–6), 1047–58. https://doi.org/10.1089/ars.2006.8.1047 (2006).
doi: 10.1089/ars.2006.8.1047 pubmed: 16771694
Moody, C. A. & Laimins, L. A. Human papillomavirus oncoproteins: Pathways to transformation. Nat. Rev. Cancer. 10(8), 550–560. https://doi.org/10.1038/nrc2886 (2010).
doi: 10.1038/nrc2886 pubmed: 20592731
Duensing, S. & Münger, K. The human papillomavirus type 16 E6 and E7 oncoproteins independently induce numerical and structural chromosome instability. Cancer Res. 62(23), 7075–7082 (2002).
pubmed: 12460929
Hochmann, J. et al. Human papillomavirus type 18 E5 oncoprotein cooperates with E6 and E7 in promoting cell viability and invasion and in modulating the cellular redox state. Mem. Inst. Oswaldo Cruz. 16(115), e190405. https://doi.org/10.1590/0074-02760190405 (2020).
doi: 10.1590/0074-02760190405
Arocena, M. et al. Using a variant of coverslip hypoxia to visualize tumor cell alterations at increasing distances from an oxygen source. J. Cell Physiol. 234(10), 16671–16678. https://doi.org/10.1002/jcp.28507 (2019).
doi: 10.1002/jcp.28507 pubmed: 30912143
D’Aiuto, N. et al. Hypoxia, acidification and oxidative stress in cells cultured at large distances from an oxygen source. Sci. Rep. 12(1), 21699. https://doi.org/10.1038/s41598-022-26205-y (2022).
doi: 10.1038/s41598-022-26205-y pubmed: 36522457 pmcid: 9755289
Williams, V. M., Filippova, M., Filippov, V., Payne, K. J. & Duerksen-Hughes, P. Human papillomavirus type 16 E6* induces oxidative stress and DNA damage. J. Virol. 88(12), 6751–61. https://doi.org/10.1128/JVI.03355-13 (2014).
doi: 10.1128/JVI.03355-13 pubmed: 24696478 pmcid: 4054338
Shim, J. H. et al. Protective effect of oxidative stress in HaCaT keratinocytes expressing E7 oncogene. Amino Acids. 34(1), 135–141. https://doi.org/10.1007/s00726-007-0499-y (2008).
doi: 10.1007/s00726-007-0499-y pubmed: 17334903
Marullo, R. et al. HPV16 E6 and E7 proteins induce a chronic oxidative stress response via NOX2 that causes genomic instability and increased susceptibility to DNA damage in head and neck cancer cells. Carcinogenesis 36(11), 1397–406. https://doi.org/10.1093/carcin/bgv126 (2015).
doi: 10.1093/carcin/bgv126 pubmed: 26354779 pmcid: 4751247
Termini, L. et al. Deregulated expression of superoxide dismutase-2 correlates with different stages of cervical neoplasia. Dis. Markers. 30(6), 275–281. https://doi.org/10.3233/DMA-2011-0784 (2011).
doi: 10.3233/DMA-2011-0784 pubmed: 21725155 pmcid: 3825245
Termini, L. et al. SOD2 immunoexpression predicts lymph node metastasis in penile cancer. BMC Clin. Pathol. 3(15), 3. https://doi.org/10.1186/s12907-015-0003-7.PMID:25745358;PMCID:PMC4350326 (2015).
doi: 10.1186/s12907-015-0003-7.PMID:25745358;PMCID:PMC4350326
Hussain, S. P., Hofseth, L. J. & Harris, C. C. Radical causes of cancer. Nat. Rev. Cancer. 3(4), 276–285. https://doi.org/10.1038/nrc1046 (2003).
doi: 10.1038/nrc1046 pubmed: 12671666
Bogdan, C., Röllinghoff, M. & Diefenbach, A. Reactive oxygen and reactive nitrogen intermediates in innate and specific immunity. Curr. Opin. Immunol. 12(1), 64–76. https://doi.org/10.1016/s0952-7915(99)00052-7 (2000).
doi: 10.1016/s0952-7915(99)00052-7 pubmed: 10679404
Fukumura, D., Kashiwagi, S. & Jain, R. K. The role of nitric oxide in tumour progression. Nat. Rev. Cancer. 6(7), 521–534. https://doi.org/10.1038/nrc1910 (2006).
doi: 10.1038/nrc1910 pubmed: 16794635
Georgescu, S. R. et al. New insights in the pathogenesis of HPV infection and the associated carcinogenic processes: The role of chronic inflammation and oxidative stress. J. Immunol. Res. 27(2018), 5315816. https://doi.org/10.1155/2018/5315816 (2018).
doi: 10.1155/2018/5315816
Carrillo-Beltrán, D. et al. Interaction between cigarette smoke and human papillomavirus 16 E6/E7 oncoproteins to induce SOD2 expression and DNA damage in head and neck cancer. Int. J. Mol. Sci. 24(8), 6907. https://doi.org/10.3390/ijms24086907 (2023).
doi: 10.3390/ijms24086907 pubmed: 37108069 pmcid: 10138975
Olcina, M. M. et al. Replication stress and chromatin context link ATM activation to a role in DNA replication. Mol. Cell. 52(5), 758–66. https://doi.org/10.1016/j.molcel.2013.10.019 (2013).
doi: 10.1016/j.molcel.2013.10.019 pubmed: 24268576 pmcid: 3898930
Economopoulou, M. et al. Histone H2AX is integral to hypoxia-driven neovascularization. Nat. Med. 15(5), 553–8. https://doi.org/10.1038/nm.1947 (2009).
doi: 10.1038/nm.1947 pubmed: 19377486 pmcid: 2713773
Collins, P. L. et al. DNA double-strand breaks induce H2Ax phosphorylation domains in a contact-dependent manner. Nat. Commun. 11(1), 3158. https://doi.org/10.1038/s41467-020-16926-x (2020).
doi: 10.1038/s41467-020-16926-x pubmed: 32572033 pmcid: 7308414
Dunkenberger, L., Reiss, K. & Del Valle, L. Comet assay for the detection of single and double-strand DNA breaks. Methods Mol. Biol. 2422, 263–269. https://doi.org/10.1007/978-1-0716-1948-3_18 (2022).
doi: 10.1007/978-1-0716-1948-3_18 pubmed: 34859412
Pu, X., Wang, Z. & Klaunig, J. E. Alkaline comet assay for assessing DNA damage in individual cells. Curr. Protoc. Toxicol. 65, 3.12.1-3.12.11. https://doi.org/10.1002/0471140856.tx0312s65 (2015).
doi: 10.1002/0471140856.tx0312s65 pubmed: 26250399
Kang, S. D. et al. Effect of productive human papillomavirus 16 infection on global gene expression in cervical epithelium. J. Virol. 92(20), e01261-e1318. https://doi.org/10.1128/JVI.01261-18 (2018).
doi: 10.1128/JVI.01261-18 pubmed: 30045992 pmcid: 6158420
Dos Santos, Á. et al. DNA damage alters nuclear mechanics through chromatin reorganization. Nucleic Acids Res. 49(1), 340–353. https://doi.org/10.1093/nar/gkaa1202 (2021).
doi: 10.1093/nar/gkaa1202 pubmed: 33330932
Kandel, M. E. et al. Phase imaging with computational specificity (PICS) for measuring dry mass changes in sub-cellular compartments. Nat. Commun. 11(1), 6256. https://doi.org/10.1038/s41467-020-20062-x (2020).
doi: 10.1038/s41467-020-20062-x pubmed: 33288761 pmcid: 7721808
Baker-Groberg, S. M. et al. Effect of ionizing radiation on the physical biology of head and neck squamous cell carcinoma cells. Cell Mol. Bioeng. 8(3), 517–525. https://doi.org/10.1007/s12195-015-0393-8 (2015).
doi: 10.1007/s12195-015-0393-8 pubmed: 26417394
Eldridge, W. J., Steelman, Z. A., Loomis, B. & Wax, A. Optical phase measurements of disorder strength link microstructure to cell stiffness. Biophys. J. 112(4), 692–702. https://doi.org/10.1016/j.bpj.2016.12.016 (2017).
doi: 10.1016/j.bpj.2016.12.016 pubmed: 28256229 pmcid: 5340128
Pérez, F. et al. Furoxans and tocopherol analogs-furoxan hybrids as anticancer agents. Anticancer Drugs 30(4), 330–338. https://doi.org/10.1097/CAD.0000000000000721 (2019).
doi: 10.1097/CAD.0000000000000721 pubmed: 30489292
Phillips, K. G., Jacques, S. L. & McCarty, O. J. Measurement of single cell refractive index, dry mass, volume, and density using a transillumination microscope. Phys. Rev. Lett. 109(11), 118105. https://doi.org/10.1103/PhysRevLett.109.118105 (2012).
doi: 10.1103/PhysRevLett.109.118105 pubmed: 23005682 pmcid: 3621783
Schmidt, U., Weigert, M., Broaddus, C., Myers, G. Cell Detection with Star-Convex Polygons. In: Frangi, A., Schnabel, J., Davatzikos, C., Alberola-López, C., Fichtinger, G. (eds) Medical Image Computing and Computer Assisted Intervention – MICCAI 2018. MICCAI 2018. Lecture Notes in Computer Science. 2018 vol 11071. Springer, Cham. https://doi.org/10.1007/978-3-030-00934-2_30
Landini G. at bham. ac. Uk BLOG. https://blog.bham.ac.uk/intellimic/g-landini - software.

Auteurs

Jimena Hochmann (J)

Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay. jhochmann@fcien.edu.uy.
Departamento de Diagnóstico en Patología y Medicina Bucal, Facultad de Odontología, Universidad de la República, General Las Heras 1925, Montevideo, Uruguay. jhochmann@fcien.edu.uy.

Magdalena Millán (M)

Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.

Paola Hernández (P)

Departamento de Genética, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.

Laura Lafon-Hughes (L)

Departamento de Genética, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.
Grupo de Biofisicoquímica, Departamento de Ciencias Biológicas, Centro Universitario Regional Litoral Norte -Sede Salto, Universidad de la República (CENUR LN, UdelaR), Montevideo, Uruguay.

Natali D' Aiuto (N)

Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.
Departamento de Biología Odontológica, Facultad de Odontología, Universidad de la República, General Las Heras 1925, Montevideo, Uruguay.

Alejandro Silva (A)

Instituto de Física, Facultad de Ingeniería, Universidad de la República, Montevideo, Uruguay.

Juan Llaguno (J)

Instituto de Física, Facultad de Ingeniería, Universidad de la República, Montevideo, Uruguay.

Julia Alonso (J)

Instituto de Física, Facultad de Ingeniería, Universidad de la República, Montevideo, Uruguay.

Ariel Fernández (A)

Instituto de Física, Facultad de Ingeniería, Universidad de la República, Montevideo, Uruguay.

Vanesa Pereira-Prado (V)

Departamento de Diagnóstico en Patología y Medicina Bucal, Facultad de Odontología, Universidad de la República, General Las Heras 1925, Montevideo, Uruguay.

José Sotelo-Silveira (J)

Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.
Sección Biología Celular, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay.

Ronell Bologna-Molina (R)

Departamento de Diagnóstico en Patología y Medicina Bucal, Facultad de Odontología, Universidad de la República, General Las Heras 1925, Montevideo, Uruguay.

Miguel Arocena (M)

Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay. m.arocena.sutz@gmail.com.
Departamento de Biología Odontológica, Facultad de Odontología, Universidad de la República, General Las Heras 1925, Montevideo, Uruguay. m.arocena.sutz@gmail.com.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Virus › Virus oncogènes › Virus tumoraux à ADN › Papillomaviridae › Virus des Papillomavirus humains › Papillomavirus humain de type 18 › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Produits chimiques inorganiques › Composés de l'oxygène › Espèces réactives de l'oxygène › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines virales › Protéines des oncogènes viraux › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Phénomènes physiologiques › Stress physiologique › Stress oxydatif › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Cellules › Cellules épithéliales › Kératinocytes › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Composants de génome › Gènes › Gènes tumoraux › Oncogènes › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › États, signes et symptômes pathologiques › Signes et symptômes › Signes et symptômes respiratoires › Hypoxie › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines tumorales › Protéines oncogènes › Protéines E7 de papillomavirus › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Maladies urogénitales › Tumeurs de l'appareil urogénital › Tumeurs de l'appareil génital féminin › Tumeurs de l'utérus › Tumeurs du col de l'utérus › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › États, signes et symptômes pathologiques › Processus pathologiques › Caractéristiques de la maladie › Maladies transmissibles › Maladies sexuellement transmissibles › Maladies sexuellement transmissibles virales › Infections à papillomavirus › Contributions of viral oncogenes of HPV-18 and...
questionsmedicales.fr › Phénomènes physiologiques cellulaires › Microenvironnement cellulaire › Microenvironnement tumoral › Contributions of viral oncogenes of HPV-18 and...