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Virology Journal

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01-12-2020 | Human Papillomavirus | Research

Human myoma tissue-based extracellular matrix models for testing the effects of irradiation on the HPV positive cells

Authors: Heidi Tuominen, Ahmed Al-Samadi, Tuula Salo, Jaana Rautava

Published in: Virology Journal | Issue 1/2020

Abstract

Background

This study was designed to investigate the invasion of human papillomavirus (HPV) positive human cervical carcinoma cell lines in human leiomyoma-based extracellular matrices in vitro, and to test the suitability of the model for studying the irradiation effects on the cancer cell invasion.

Methods

HPV positive cervical carcinoma cell lines SiHa and CaSki, and HPV negative squamous cell carcinoma cell line HSC-3 were used. CaSki cells contain around 600 copies of HPV 16 virus in the genome, whereas SiHa have only 1–2 copies per cell. Cells were analyzed using two different human tumor derived extracellular matrix methods (3D myoma disc model, and Myogel Transwell invasion assay). Cultures were irradiated with 4 Gy. Myoma invasion area and the depth of invasion were measured with ImageJ 1.51j8 software. Statistical analyses were performed with SPSS Statistics (IBM SPSS® Statistics 25).

Results

All cells invaded through Myogel coated Transwell membranes and within myoma discs. In myoma discs, a difference in the invasion depth (p = 0.0001) but not in invasion area (p = 0.310) between the HPV positive cell lines was seen, since SiHa (less HPV) invaded slightly better than CaSki (more HPV). HSC-3 cells (HPV negative) invaded deepest (p = 0.048) than either of the HPV positive cell line cells. No difference was detected in the invasion area (p = 0.892) between HPV positive and HPV negative cells. The ionized radiation significantly reduced the invasion depth of HSC-3 (p = 0.008), SiHa (p = 0.0001) and CaSki (p = 0.005). No significant effect on the invasion area was detected in any of the cell lines. However, a significant difference was observed between SiHa and CaSki in the reduction of the invasion depth after radiation (p = 0.013) as the reduction was greater with SiHa than CaSki.

Conclusions

Both solid and gelatinous human leiomyoma-based extracellular matrix models were suitable platforms to study the invasion of HPV positive cervical carcinoma cells in vitro. SiHa cells with less HPV copy number cells invaded slightly better and were slightly more sensitive to irradiation than CaSki cells with high HPV copy number. However, there was no drastic differences between the invasion properties of these carcinoma cells.

Salo T, Dourado MR, Sundquist E, et al. Organotypic three-dimensional assays based on human leiomyoma–derived matrices. Philos Trans R Soc B Biol Sci. 2018;373(1737):20160482. https://doi.org/10.1098/rstb.2016.0482.CrossRef

Sundquist E, Renko O, Salo S, et al. Neoplastic extracellular matrix environment promotes cancer invasion in vitro. Exp Cell Res. 2016;344(2):229–40. https://doi.org/10.1016/j.yexcr.2016.04.003.CrossRefPubMed

Salo T, Sutinen M, Hoque Apu E, et al. A novel human leiomyoma tissue derived matrix for cell culture studies. BMC Cancer. 2015;15(1):981. https://doi.org/10.1186/s12885-015-1944-z.CrossRefPubMedPubMedCentral

Nurmenniemi S, Sinikumpu T, Alahuhta I, et al. A novel organotypic model mimics the tumor microenvironment. Am J Pathol. 2009;175(3):1281–91. https://doi.org/10.2353/ajpath.2009.081110.CrossRefPubMedPubMedCentral

Åström P, Heljasvaara R, Nyberg P, Al-Samadi A, Salo T. Human tumor tissue-based 3D in vitro invasion assays. Methods Mol Biol (Clifton, N.J.). 2018;1731:213–21. https://doi.org/10.1007/978-1-4939-7595-2_19.CrossRef

Naakka E, Tuomainen K, Wistrand H, et al. Fully human tumor-based matrix in three-dimensional spheroid invasion assay. J Vis Exp. 2019;(147). https://doi.org/10.3791/59567.

Al-Samadi A, Awad SA, Tuomainen K, et al. Crosstalk between tongue carcinoma cells, extracellular vesicles, and immune cells in in vitro and in vivo models. Oncotarget. 2017;8(36):60123–34. https://doi.org/10.18632/oncotarget.17768.CrossRefPubMedPubMedCentral

Teppo S, Sundquist E, Vered M, et al. The hypoxic tumor microenvironment regulates invasion of aggressive oral carcinoma cells. Exp Cell Res. 2013;319(4):376–89. https://doi.org/10.1016/j.yexcr.2012.12.010.CrossRefPubMed

Alahuhta I, Aikio M, Väyrynen O, et al. Endostatin induces proliferation of oral carcinoma cells but its effect on invasion is modified by the tumor microenvironment. Exp Cell Res. 2015;336(1):130–40. https://doi.org/10.1016/j.yexcr.2015.06.012.CrossRefPubMed

10.

Dayan D, Salo T, Salo S, et al. Molecular crosstalk between cancer cells and tumor microenvironment components suggests potential targets for new therapeutic approaches in mobile tongue cancer. Cancer Med. 2012;1(2):128–40. https://doi.org/10.1002/cam4.24.CrossRefPubMedPubMedCentral

11.

Pirilä E, Väyrynen O, Sundquist E, et al. Macrophages modulate migration and invasion of human tongue squamous cell carcinoma. Guan X-Y, ed. PLoS One. 2015;10(3):e0120895. https://doi.org/10.1371/journal.pone.0120895.CrossRefPubMedPubMedCentral

12.

Korvala J, Jee K, Porkola E, et al. MicroRNA and protein profiles in invasive versus non-invasive oral tongue squamous cell carcinoma cells in vitro. Exp Cell Res. 2017;350(1):9–18. https://doi.org/10.1016/j.yexcr.2016.10.015.CrossRefPubMed

13.

Väyrynen O, Åström P, Nyberg P, et al. Matrix metalloproteinase 9 inhibits the motility of highly aggressive HSC-3 oral squamous cell carcinoma cells. Exp Cell Res. 2019;376(1):18–26. https://doi.org/10.1016/j.yexcr.2019.01.018.CrossRefPubMed

14.

Kauppila JH, Korvala J, Siirilä K, et al. Toll-like receptor 9 mediates invasion and predicts prognosis in squamous cell carcinoma of the mobile tongue. J Oral Pathol Med. 2015;44(8):571–7. https://doi.org/10.1111/jop.12272.CrossRefPubMed

15.

Mäkinen LK, Ahmed A, Hagström J, et al. Toll-like receptors 2, 4, and 9 in primary, metastasized, and recurrent oral tongue squamous cell carcinomas. J Oral Pathol Med. 2016;45(5):338–45. https://doi.org/10.1111/jop.12373.CrossRefPubMed

16.

Giuliano AR, Nyitray AG, Kreimer AR, et al. EUROGIN 2014 roadmap: differences in HPV infection natural history, transmission, and HPV-related cancer incidence by gender and anatomic site of infection HHS public access. Int J Cancer Int J Cancer. 2015. https://doi.org/10.1002/ijc.29082.

17.

Yee C, Krishnan-Hewlett I, Baker CC, Schlegel R, Howley PM. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am J Pathol. 1985;119(3):361–6 http://www.ncbi.nlm.nih.gov/pubmed/2990217. Accessed 20 May 2019.PubMedPubMedCentral

18.

Raybould R, Fiander A, Wilkinson GWG, Hibbitts S. HPV integration detection in CaSki and SiHa using detection of integrated papillomavirus sequences and restriction-site PCR. J Virol Methods. 2014;206:51–4. https://doi.org/10.1016/j.jviromet.2014.05.017.CrossRefPubMed

19.

Pater M, Pater A. Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix. Virology. 1985;145(2):313–8.CrossRefPubMed

20.

Ang KK, Harris J, Wheeler R, et al. Human papillomavirus and survival of patients with oropharyngeal cancer. N Engl J Med. 2010;363(1):24–35. https://doi.org/10.1056/NEJMoa0912217.CrossRefPubMedPubMedCentral

21.

Kimple RJ, Smith MA, Blitzer GC, et al. Enhanced radiation sensitivity in HPV-positive head and neck cancer. Cancer Res. 2013;73(15):4791–800. https://doi.org/10.1158/0008-5472.CAN-13-0587.CrossRefPubMedPubMedCentral

22.

Zhang M, Rose B, Lee CS, Hong AM. In vitro 3-dimensional tumor model for radiosensitivity of HPV positive OSCC cell lines. Cancer Biol Ther. 2015;16(8):1231–40. https://doi.org/10.1080/15384047.2015.1056410.CrossRefPubMedPubMedCentral

23.

Ibragimova M, Tsyganov M, Shpileva O, et al. HPV status and its genomic integration affect survival of patients with cervical cancer. Neoplasma. 2018;65(3):441–8. https://doi.org/10.4149/neo_2018_170414N277.CrossRefPubMed

24.

Bachtiary B, Obermair A, Dreier B, et al. Impact of multiple HPV infection on response to treatment and survival in patients receiving radical radiotherapy for cervical cancer. Int J Cancer. 2002;102(3):237–43. https://doi.org/10.1002/ijc.10708.CrossRefPubMed

25.

Harima Y, Sawada S, Nagata K, Sougawa M, Ohnishi T. Human papilloma virus (HPV) DNA associat human papilloma virus (HPV) DNA associated with prognosis of cervical cancer after radiotherapy. Int J Radiat Oncol Biol Phys. 2002;52(5):1345–51. https://doi.org/10.1016/S0360-3016(01)02796-1.CrossRefPubMed

26.

Li P, Tan Y, Zhu LX, et al. Prognostic value of HPV DNA status in cervical cancer before treatment: a systematic review and meta-analysis. Oncotarget. 2017;8(39):66352–9. https://doi.org/10.18632/oncotarget.18558.CrossRefPubMedPubMedCentral

27.

Riou G, Favre M, Jeannel D, Bourhis J, Le Doussal V, Orth G. Association between poor prognosis in early-stage invasive cervical carcinomas and non-detection of HPV DNA. Lancet (London, England). 1990;335(8699):1171–4. https://doi.org/10.1016/0140-6736(90)92693-c.CrossRef

28.

Crook T, Vousden KH. Properties of p53 mutations detected in primary and secondary cervical cancers suggest mechanisms of metastasis and involvement of environmental carcinogens. EMBO J. 1992;11(11):3935–40 http://www.ncbi.nlm.nih.gov/pubmed/1327751. Accessed 12 Oct 2019.CrossRefPubMedPubMedCentral

29.

Kibbey MC. Maintenance of the EHS sarcoma and Matrigel preparation. J Tissue Cult Methods. 1994;16(3–4):227–30. https://doi.org/10.1007/BF01540656.CrossRef

30.

Väyrynen O, Piippo M, Jämsä H, et al. Effects of ionizing radiation and HPSE1 inhibition on the invasion of oral tongue carcinoma cells on human extracellular matrices in vitro. Exp Cell Res. 2018;371(1):151–61. https://doi.org/10.1016/j.yexcr.2018.08.005.CrossRefPubMed

31.

van Wyk HC, Roseweir A, Alexander P, et al. The relationship between tumor budding, tumor microenvironment, and survival in patients with primary operable colorectal cancer. Ann Surg Oncol. 2019. https://doi.org/10.1245/s10434-019-07931-6.

32.

Mäkitie AA, Almangush A, Rodrigo JP, Ferlito A, Leivo I. Hallmarks of cancer: tumor budding as a sign of invasion and metastasis in head and neck cancer. Head Neck. 2019;41(10):3712–8. https://doi.org/10.1002/hed.25872.CrossRefPubMed

33.

Seki M, Sano T, Yokoo S, Oyama T. Tumour budding evaluated in biopsy specimens is a useful predictor of prognosis in patients with cN0 early stage oral squamous cell carcinoma. Histopathology. 2017;70(6):869–79. https://doi.org/10.1111/his.13144.CrossRefPubMed

34.

Almangush A, Pirinen M, Heikkinen I, Mäkitie AA, Salo T, Leivo I. Tumour budding in oral squamous cell carcinoma: a meta-analysis. Br J Cancer. 2018;118(4):577–86. https://doi.org/10.1038/bjc.2017.425.CrossRefPubMed

35.

Almangush A, Leivo I, Siponen M, et al. Evaluation of the budding and depth of invasion (BD) model in oral tongue cancer biopsies. Virchows Arch. 2018;472(2):231–6. https://doi.org/10.1007/s00428-017-2212-1.CrossRefPubMed

36.

Almangush A, Karhunen M, Hautaniemi S, Salo T, Leivo I. Prognostic value of tumour budding in oesophageal cancer: a meta-analysis. Histopathology. 2016;68(2):173–82. https://doi.org/10.1111/his.12781.CrossRefPubMed

37.

Chatterjee D, Bansal V, Malik V, et al. Tumor budding and worse pattern of invasion can predict nodal metastasis in oral cancers and associated with poor survival in early-stage tumors. Ear Nose Throat J. 2019;98(7):E112–9. https://doi.org/10.1177/0145561319848669.CrossRefPubMed

38.

Ebihara Y, Yoshida S, Nakahira M, et al. Importance of tumor budding grade as independent prognostic factor for early tongue squamous cell carcinoma. Head Neck. 2019;41(6):1809–15. https://doi.org/10.1002/hed.25614.CrossRefPubMed

39.

Angadi PV, Patil PV, Hallikeri K, Mallapur MD, Hallikerimath S, Kale AD. Tumor budding is an independent prognostic factor for prediction of lymph node metastasis in oral squamous cell carcinoma. Int J Surg Pathol. 2015;23(2):102–10. https://doi.org/10.1177/1066896914565022.CrossRefPubMed

40.

Zhu Y, Liu H, Xie N, et al. Impact of tumor budding in head and neck squamous cell carcinoma: a meta-analysis. Head Neck. 2019;41(2):542–50. https://doi.org/10.1002/hed.25462.CrossRefPubMed

41.

Satabongkoch N, Khunamornpong S, Pongsuvareeyakul T, et al. Prognostic value of tumor budding in early-stage cervical adenocarcinomas. Asian Pac J Cancer Prev. 2017;18(6):1717–22. https://doi.org/10.22034/APJCP.2017.18.6.1717.CrossRefPubMedPubMedCentral

42.

Jesinghaus M, Strehl J, Boxberg M, et al. Introducing a novel highly prognostic grading scheme based on tumour budding and cell nest size for squamous cell carcinoma of the uterine cervix. J Pathol Clin Res. 2018;4(2):93–102. https://doi.org/10.1002/cjp2.95.CrossRefPubMedPubMedCentral

43.

Wang MB, Liu IY, Gornbein JA, Nguyen CT. HPV-positive oropharyngeal carcinoma. Otolaryngol Neck Surg. 2015;153(5):758–69. https://doi.org/10.1177/0194599815592157.CrossRef

44.

Nagel R, Martens-de Kemp SR, Buijze M, Jacobs G, Braakhuis BJM, Brakenhoff RH. Treatment response of HPV-positive and HPV-negative head and neck squamous cell carcinoma cell lines. Oral Oncol. 2013;49(6):560–6. https://doi.org/10.1016/j.oraloncology.2013.03.446.CrossRefPubMed

45.

Schneider K, Bol V, Grégoire V. Lack of differences in radiation-induced immunogenicity parameters between HPV-positive and HPV-negative human HNSCC cell lines. Radiother Oncol. 2017;124(3):411–7. https://doi.org/10.1016/j.radonc.2017.08.018.CrossRefPubMed

46.

Sørensen BS, Busk M, Olthof N, et al. Radiosensitivity and effect of hypoxia in HPV positive head and neck cancer cells. Radiother Oncol. 2013;108(3):500–5. https://doi.org/10.1016/j.radonc.2013.06.011.CrossRefPubMed

Title: Human myoma tissue-based extracellular matrix models for testing the effects of irradiation on the HPV positive cells
Authors: Heidi Tuominen
Ahmed Al-Samadi
Tuula Salo
Jaana Rautava
Publication date: 01-12-2020
Publisher: BioMed Central
Keywords: Human Papillomavirus
Cervical Cancer
Cervical Cancer
Published in: Virology Journal / Issue 1/2020
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-020-01367-1

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Human myoma tissue-based extracellular matrix models for testing the effects of irradiation on the HPV positive cells

Abstract

Background

Methods

Results

Conclusions

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Abstract

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Conclusions

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