1887

Journal of General Virology header logo

Volume 99, Issue 9

Impact of human cytomegalovirus on glioblastoma cell viability and chemotherapy treatment Free

Abstract

The relationship between human cytomegalovirus (HCMV) and tumours has been extensively investigated, mainly in glioblastoma multiforme (GBM), a malignant tumour of the central nervous system with low overall survival rates. Several reports have demonstrated the presence of HCMV in GBM, although typically restricted to a low number of cells, and studies have indicated that viral proteins have the ability to dysregulate cellular processes and increase tumour malignancy. Treatment of GBM involves the use of the chemotherapeutic agents temozolomide (TMZ) and carmustine (bis-chloroethylnitrosourea, BCNU), which lead to the attachment of adducts to the DNA backbone, causing errors during replication and consequent cell death. It is known that HCMV infection can modulate DNA repair pathways, but what effects the virus may exhibit during chemotherapy are unknown. Here we approach this question by analysing HCMV infection and viral protein accumulation in GBM cell lines with different genotypes and their response to TMZ and BCNU in the presence of the virus. We demonstrate that A172, TP365MG and U251MG GBM cells are efficiently infected by both low-passage (TB40E) and high-passage (AD169) HCMV strains. However, the GBM cell lines vary widely in their permissiveness to viral gene expression and exhibit very different patterns of immediate early, early and late protein accumulation. HCMV reduces the viability of permissive GBM cells in a multiplicity-dependent manner in both the absence and presence of TMZ or BNCU. In sum, we demonstrate that GBM cell lines are equally susceptible but differentially permissive to infection by both low- and high-passage strains of HCMV. This observation not only indicates that viral replication is largely controlled by cellular factors in this system, but also provides a possible explanation for why viral gene products are only found in a subset of cells in GBM tumours. Furthermore, we conclude that the virus does not confer increased resistance to chemotherapeutic drugs in various GBM cell lines, but instead reduces tumour cell viability. These results highlight that the oncomodulatory potential of HCMV is not limited to cancer-promoting activities, but also includes adverse effects on tumour cell proliferation or survival.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): carmustina , chemotherapy , glioblastoma multiforme , human cytomegalovirus and temozolomide
© 2018 The Authors
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001118
2018-07-27
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/99/9/1274.html?itemId=/content/journal/jgv/10.1099/jgv.0.001118&mimeType=html&fmt=ahah

References

  1. Griffiths P, Baraniak I, Reeves M. The pathogenesis of human cytomegalovirus. J Pathol 2015; 235:288–297 [View Article][PubMed]
     [Google Scholar]
  2. Nogalski MT, Collins-McMillen D, Yurochko AD. Overview of human cytomegalovirus pathogenesis. Methods Mol Biol 2014; 1119:15–28 [View Article][PubMed]
     [Google Scholar]
  3. Banerjee S, Wei Z, Tan F, Peck KN, Shih N et al. Distinct microbiological signatures associated with triple negative breast cancer. Sci Rep 2015; 5:15162 [View Article][PubMed]
     [Google Scholar]
  4. Baryawno N, Rahbar A, Wolmer-Solberg N, Taher C, Odeberg J et al. Detection of human cytomegalovirus in medulloblastomas reveals a potential therapeutic target. J Clin Invest 2011; 121:4043–4055 [View Article]
     [Google Scholar]
  5. Bhattacharjee B, Renzette N, Kowalik TF. Genetic analysis of cytomegalovirus in malignant gliomas. J Virol 2012; 86:6815–6824 [View Article][PubMed]
     [Google Scholar]
  6. Harkins LE, Matlaf LA, Soroceanu L, Klemm K, Britt WJ et al. Detection of human cytomegalovirus in normal and neoplastic breast epithelium. Herpesviridae 2010; 1:8 [View Article][PubMed]
     [Google Scholar]
  7. Samanta M, Harkins L, Klemm K, Britt WJ, Cobbs CS. High prevalence of human cytomegalovirus in prostatic intraepithelial neoplasia and prostatic carcinoma. J Urol 2003; 170:998–1002 [View Article][PubMed]
     [Google Scholar]
  8. Taher C, de Boniface J, Mohammad AA, Religa P, Hartman J et al. High prevalence of human cytomegalovirus proteins and nucleic acids in primary breast cancer and metastatic sentinel lymph nodes. PLoS One 2013; 8:e56795 [View Article][PubMed]
     [Google Scholar]
  9. Price RL, Harkins L, Chiocca EA, Zhang PJ, Kurt H et al. Human cytomegalovirus is present in alveolar soft part sarcoma. Appl Immunohistochem Mol Morphol 2017; 25:615–619 [View Article][PubMed]
     [Google Scholar]
  10. Cui J, Wang Q, Wang HB, Wang B, Li L. Protein and DNA evidences of HCMV infection in primary breast cancer tissues and metastatic sentinel lymph nodes. Cancer Biomark 2018; 21:769–780 [View Article][PubMed]
     [Google Scholar]
  11. Carlson JW, Rådestad AF, Söderberg-Naucler C, Rahbar A. Human cytomegalovirus in high grade serous ovarian cancer possible implications for patients survival. Medicine 2018; 97:e9685 [View Article][PubMed]
     [Google Scholar]
  12. Bai B, Wang X, Chen E, Zhu H. Human cytomegalovirus infection and colorectal cancer risk: a meta-analysis. Oncotarget 2016; 7:76735–76742 [View Article][PubMed]
     [Google Scholar]
  13. Cobbs CS, Harkins L, Samanta M, Gillespie GY, Bharara S et al. Human cytomegalovirus infection and expression in human malignant glioma. Cancer Res 2002; 62:3347–3350[PubMed]
     [Google Scholar]
  14. Stangherlin LM, Castro FL, Medeiros RS, Guerra JM, Kimura LM et al. Human cytomegalovirus DNA quantification and gene expression in gliomas of different grades. PLoS One 2016; 11:e0159604 [View Article][PubMed]
     [Google Scholar]
  15. dos Santos CJ, Stangherlin LM, Figueiredo EG, Corrêa C, Teixeira MJ et al. High prevalence of HCMV and viral load in tumor tissues and peripheral blood of glioblastoma multiforme patients. J Med Virol 2014; 86:1953–1961 [View Article][PubMed]
     [Google Scholar]
  16. McFaline-Figueroa JR, Wen PY. The viral connection to glioblastoma. Curr Infect Dis Rep 2017; 19:5 [View Article][PubMed]
     [Google Scholar]
  17. Wen PY, Kesari S. Malignant gliomas in adults. N Engl J Med 2008; 359:492–507 [View Article][PubMed]
     [Google Scholar]
  18. Ramirez YP, Weatherbee JL, Wheelhouse RT, Ross AH. Glioblastoma multiforme therapy and mechanisms of resistance. Pharmaceuticals 2013; 6:1475–1506 [View Article][PubMed]
     [Google Scholar]
  19. Cobbs CS, Soroceanu L, Denham S, Zhang W, Kraus MH. Modulation of oncogenic phenotype in human glioma cells by cytomegalovirus IE1-mediated mitogenicity. Cancer Res 2008; 68:724–730 [View Article][PubMed]
     [Google Scholar]
  20. Strååt K, Liu C, Rahbar A, Zhu Q, Liu L et al. Activation of telomerase by human cytomegalovirus. J Natl Cancer Inst 2009; 101:488–497 [View Article][PubMed]
     [Google Scholar]
  21. Cobbs CS, Soroceanu L, Denham S, Zhang W, Britt WJ et al. Human cytomegalovirus induces cellular tyrosine kinase signaling and promotes glioma cell invasiveness. J Neurooncol 2007; 85:271–280 [View Article][PubMed]
     [Google Scholar]
  22. Soroceanu L, Matlaf L, Bezrookove V, Harkins L, Martinez R et al. Human cytomegalovirus US28 found in glioblastoma promotes an invasive and angiogenic phenotype. Cancer Res 2011; 71:6643–6653 [View Article][PubMed]
     [Google Scholar]
  23. Fornara O, Bartek J, Rahbar A, Odeberg J, Khan Z et al. Cytomegalovirus infection induces a stem cell phenotype in human primary glioblastoma cells: prognostic significance and biological impact. Cell Death Differ 2016; 23:261–269 [View Article][PubMed]
     [Google Scholar]
  24. Fiallos E, Judkins J, Matlaf L, Prichard M, Dittmer D et al. Human cytomegalovirus gene expression in long-term infected glioma stem cells. PLoS One 2014; 9:e116178 [View Article][PubMed]
     [Google Scholar]
  25. Maussang D, Verzijl D, van Walsum M, Leurs R, Holl J et al. Human cytomegalovirus-encoded chemokine receptor US28 promotes tumorigenesis. Proc Natl Acad Sci USA 2006; 103:13068–13073 [View Article][PubMed]
     [Google Scholar]
  26. Slinger E, Maussang D, Schreiber A, Siderius M, Rahbar A et al. HCMV-encoded chemokine receptor US28 mediates proliferative signaling through the IL-6-STAT3 axis. Sci Signal 2010; 3:ra58 [View Article][PubMed]
     [Google Scholar]
  27. Matlaf LA, Harkins LE, Bezrookove V, Cobbs CS, Soroceanu L. Cytomegalovirus pp71 protein is expressed in human glioblastoma and promotes pro-angiogenic signaling by activation of stem cell factor. PLoS One 2013; 8:e68176 [View Article][PubMed]
     [Google Scholar]
  28. Dziurzynski K, Wei J, Qiao W, Hatiboglu MA, Kong LY et al. Glioma-associated cytomegalovirus mediates subversion of the monocyte lineage to a tumor propagating phenotype. Clin Cancer Res 2011; 17:4642–4649 [View Article][PubMed]
     [Google Scholar]
  29. Cinatl J, Scholz M, Doerr HW. Role of tumor cell immune escape mechanisms in cytomegalovirus-mediated oncomodulation. Med Res Rev 2005; 25:167–185 [View Article][PubMed]
     [Google Scholar]
  30. Foster H, Ulasov IV, Cobbs CS. Human cytomegalovirus-mediated immunomodulation: Effects on glioblastoma progression. Biochim Biophys Acta 2017; 1868:273–276 [View Article][PubMed]
     [Google Scholar]
  31. Joseph GP, McDermott R, Baryshnikova MA, Cobbs CS, Ulasov IV. Cytomegalovirus as an oncomodulatory agent in the progression of glioma. Cancer Lett 2017; 384:79–85 [View Article][PubMed]
     [Google Scholar]
  32. Batista LF, Roos WP, Christmann M, Menck CF, Kaina B. Differential sensitivity of malignant glioma cells to methylating and chloroethylating anticancer drugs: p53 determines the switch by regulating xpc, ddb2, and DNA double-strand breaks. Cancer Res 2007; 67:11886–11895 [View Article][PubMed]
     [Google Scholar]
  33. Li QJ, Cai JQ, Liu CY. Evolving molecular genetics of glioblastoma. Chin Med J 2016; 129:464–471 [View Article][PubMed]
     [Google Scholar]
  34. Erasimus H, Gobin M, Niclou S, van Dyck E. DNA repair mechanisms and their clinical impact in glioblastoma. Mutat Res Rev Mutat Res 2016; 769:19–35 [View Article][PubMed]
     [Google Scholar]
  35. Ranneberg-Nilsen T, Bjørås M, Luna L, Slettebakk R, Dale HA et al. Human cytomegalovirus infection modulates DNA base excision repair in fibroblast cells. Virology 2006; 348:389–397 [View Article][PubMed]
     [Google Scholar]
  36. Costa H, Nascimento R, Sinclair J, Parkhouse RM. Human cytomegalovirus gene UL76 induces IL-8 expression through activation of the DNA damage response. PLoS Pathog 2013; 9:e1003609 [View Article][PubMed]
     [Google Scholar]
  37. O'Dowd JM, Zavala AG, Brown CJ, Mori T, Fortunato EA. HCMV-infected cells maintain efficient nucleotide excision repair of the viral genome while abrogating repair of the host genome. PLoS Pathog 2012; 8:e1003038 [View Article][PubMed]
     [Google Scholar]
  38. Siew VK, Duh CY, Wang SK. Human cytomegalovirus UL76 induces chromosome aberrations. J Biomed Sci 2009; 16:107 [View Article][PubMed]
     [Google Scholar]
  39. Fortunato EA, dell'aquila ML, Spector DH. Specific chromosome 1 breaks induced by human cytomegalovirus. Proc Natl Acad Sci USA 2000; 97:853–858 [View Article][PubMed]
     [Google Scholar]
  40. Nystad M, Fagerheim T, Brox V, Fortunato EA, Nilssen Ø. Human cytomegalovirus (HCMV) and hearing impairment: infection of fibroblast cells with HCMV induces chromosome breaks at 1q23.3, between loci DFNA7 and DFNA49 – both involved in dominantly inherited, sensorineural, hearing impairment. Mutat Res 2008; 637:56–65 [View Article][PubMed]
     [Google Scholar]
  41. Korbecki J, Gutowska I, Kojder I, Jeżewski D, Goschorska M et al. New extracellular factors in glioblastoma multiforme development: neurotensin, growth differentiation factor-15, sphingosine-1-phosphate and cytomegalovirus infection. Oncotarget 2018; 9:7219–7270 [View Article][PubMed]
     [Google Scholar]
  42. Zhang X, Liu P, Zhang B, Wang A, Yang M. Role of STAT3 decoy oligodeoxynucleotides on cell invasion and chemosensitivity in human epithelial ovarian cancer cells. Cancer Genet Cytogenet 2010; 197:46–53 [View Article][PubMed]
     [Google Scholar]
  43. Reinartz R, Wang S, Kebir S, Silver DJ, Wieland A et al. Functional subclone profiling for prediction of treatment-induced intratumor population shifts and discovery of rational drug combinations in human glioblastoma. Clin Cancer Res 2017; 23:562–574 [View Article][PubMed]
     [Google Scholar]
  44. Lau SK, Chen YY, Chen WG, Diamond DJ, Mamelak AN et al. Lack of association of cytomegalovirus with human brain tumors. Mod Pathol 2005; 18:838–843 [View Article][PubMed]
     [Google Scholar]
  45. Taha MS, Abdalhamid BA, El-Badawy SA, Sorour YM, Almsned FM et al. Expression of cytomegalovirus in glioblastoma multiforme: Myth or reality?. Br J Neurosurg 2016; 30:307–312 [View Article][PubMed]
     [Google Scholar]
  46. Garcia-Martinez A, Alenda C, Irles E, Ochoa E, Quintanar T et al. Lack of cytomegalovirus detection in human glioma. Virol J 2017; 14:216 [View Article][PubMed]
     [Google Scholar]
  47. American Type Culture Collection 2017; ATCC tumor cell panel. www.atcc.org/~/media/PDFs/Culture Guides/TumorCellPanelsBrochure.ashx
  48. Bouaoun L, Sonkin D, Ardin M, Hollstein M, Byrnes G et al. TP53 variations in human cancers: new lessons from the IARC TP53 database and genomics data. Hum Mutat 2016; 37:865–876 [View Article][PubMed]
     [Google Scholar]
  49. Ishii N, Maier D, Merlo A, Tada M, Sawamura Y et al. Frequent co-alterations of TP53, p16/CDKN2A, p14ARF, PTEN tumor suppressor genes in human glioma cell lines. Brain Pathol 1999; 9:469–479 [View Article][PubMed]
     [Google Scholar]
  50. Murphy ÁC, Weyhenmeyer B, Schmid J, Kilbride SM, Rehm M et al. Activation of executioner caspases is a predictor of progression-free survival in glioblastoma patients: a systems medicine approach. Cell Death Dis 2013; 4:e629 [View Article][PubMed]
     [Google Scholar]
  51. Batista LFZ, Roos WP, Kaina B, Batista LFZ, Roos WP et al. p53 mutant human glioma cells are sensitive to UV-C-induced apoptosis due to impaired cyclobutane pyrimidine dimer removal p53 mutant human glioma cells are sensitive to UV-C-induced apoptosis due to impaired cyclobutane pyrimidine dimer removal; 2009; 7237–247
  52. Lee SY. Temozolomide resistance in glioblastoma multiforme. Genes Dis 2016; 3:198–210 [View Article]
     [Google Scholar]
  53. Sijmons S, van Ranst M, Maes P. Genomic and functional characteristics of human cytomegalovirus revealed by next-generation sequencing. Viruses 2014; 6:1049–1072 [View Article][PubMed]
     [Google Scholar]
  54. Terrasson J, Allart S, Martin H, Lulé J, Haddada H et al. p73-dependent apoptosis through death receptor: impairment by human cytomegalovirus infection. Cancer Res 2005; 65:2787–2794 [View Article][PubMed]
     [Google Scholar]
  55. Allart S, Martin H, Detraves C, Terrasson J, Caput D et al. Human cytomegalovirus induces drug resistance and alteration of programmed cell death by accumulation of deltaN-p73alpha. J Biol Chem 2002; 277:29063–29068 [View Article][PubMed]
     [Google Scholar]
  56. Kim J, Kwon YJ, Park ES, Sung B, Kim JH et al. Human cytomegalovirus (HCMV) IE1 plays role in resistance to apoptosis with etoposide in cancer cell line by Cdk2 accumulation. Microbiol Immunol 2003; 47:959–967 [View Article][PubMed]
     [Google Scholar]
  57. Valle Oseguera CA, Spencer JV. cmvIL-10 stimulates the invasive potential of MDA-MB-231 breast cancer cells. PLoS One 2014; 9:e88708 [View Article][PubMed]
     [Google Scholar]
  58. Cinatl J, Cinatl J, Vogel JU, Kotchetkov R, Driever PH et al. Persistent human cytomegalovirus infection induces drug resistance and alteration of programmed cell death in human neuroblastoma cells. Cancer Res 1998; 58:367–372[PubMed]
     [Google Scholar]
  59. Liu C, Clark PA, Kuo JS, Kalejta RF. Human cytomegalovirus-infected glioblastoma cells display stem cell-like phenotypes. mSphere 2017; 2:e00137-17 [View Article][PubMed]
     [Google Scholar]
  60. Albrecht T, Deng CZ, Abdel-Rahman SZ, Fons M, Cinciripini P et al. Differential mutagen sensitivity of peripheral blood lymphocytes from smokers and nonsmokers: effect of human cytomegalovirus infection. Environ Mol Mutagen 2004; 43:169–178 [View Article][PubMed]
     [Google Scholar]
  61. Deng CZ, Abubakar S, Fons MP, Boldogh I, Hokanson J et al. Cytomegalovirus-enhanced induction of chromosome aberrations in human peripheral blood lymphocytes treated with potent genotoxic agents. Environ Mol Mutagen 1992; 19:304–310 [View Article][PubMed]
     [Google Scholar]
  62. Tanaka K, Zou JP, Takeda K, Ferrans VJ, Sandford GR et al. Effects of human cytomegalovirus immediate-early proteins on p53-mediated apoptosis in coronary artery smooth muscle cells. Circulation 1999; 99:1656–1659 [View Article][PubMed]
     [Google Scholar]
  63. Moorman NJ, Sharon-Friling R, Shenk T, Cristea IM. A targeted spatial-temporal proteomics approach implicates multiple cellular trafficking pathways in human cytomegalovirus virion maturation. Mol Cell Proteomics 2010; 9:851–860 [View Article][PubMed]
     [Google Scholar]
  64. Zhu H, Shen Y, Shenk T. Human cytomegalovirus IE1 and IE2 proteins block apoptosis. J Virol 1995; 69:7960–7970[PubMed]
     [Google Scholar]
  65. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65:55–63 [View Article][PubMed]
     [Google Scholar]
  66. Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 1986; 89:271–277[PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001118
Loading
Impact of human cytomegalovirus on glioblastoma cell viability and chemotherapy treatment
J. Gen. Virol. 99, 1274 (2018); https://doi.org/10.1099/jgv.0.001118
/content/journal/jgv/10.1099/jgv.0.001118
/content/journal/jgv/10.1099/jgv.0.001118
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error