The Association Between Tumor Necrosis Factor-Alpha (Tnf-Α) Expression and Molecular Subtype in Oral Cancer

Yıl 2020, , 1 - 7, 28.09.2020

İpek Atak Seçen , Leyla Bozdağ , Reinhard Buettner Sibel Elif Gültekin

https://doi.org/10.20515/otd.770386

Öz

Oral squamous cell carcinoma (OSCC) is the most common malignant tumor of the oral cavity and the development and progression of this tumor is due to the accumulation of genetic alterations. On the other hand, tumor necrosis factor alpha (TNF-α) is known to play an important role in tumor invasion. The aim of this study was to detect genetic alterations in oral cancer and to investigate the possible relationship between molecular subtype, inflammation and TNF-α expression. The study was conducted on 39 tissue samples, 25 of which were diagnosed as OSCC. All were subjecte to DNA isolation from paraffin embedded blocks and followed by next-generation sequencing (NGS) platform that examined the complete coding sequence of in 28 cancer-related genes. TNF-α expression in tissues was determined by immunohistochemical studies. Inflammation scoring was performed in hemotoxylin & eosin sections for each case. Eighteen of 25 (72%) OSCC cases showed alterations in at least in one gene. P53 was the most frequent altered gene in all lesions. Three or more gene alterations were detected in 3 of all cases (%12). TNF-α was overexpressed in tumors with mutation. The mutation spectrum revealed by the next-generation sequence analysis (NGS) suggests that genetic alterations in OSCC are diverse and complex. The results of the study suggest that there is a relationship between genetic alterations and inflammation/TNF-α expression.

Anahtar Kelimeler

Next-generation sequence analysis, NGS, oral cancer, TNF-α

Oral Kanserde Tümör Nekrozis Faktör-Alfa (Tnf-α) Ekspresyonunun Moleküler Subtipleme ile İlişkisi

Öz

Oral kavitenin en sık izlenen malign tümörü olan oral skuamöz hücreli karsinomun (OSHK) gelişmesi ve ilerlemesi, genetik alterasyonların birikmesinden kaynaklanmaktadır. Diğer taraftan Tümör nekrozis faktör-alfa (TNF-α)'nın tümör invazyonunda önemli rol oynadığı da bilinmektedir. Bu çalışmanın amacı, oral kanserdeki genetik alterasyonları saptayıp, moleküler alttipleme ile inflamasyon ve TNF- α ekspresyonu arasındaki ilişkiyi incelemektir. Çalışma, 25’i OSHK tanısı almış toplam 39 doku örneği üzerinde gerçekleştirildi. Parafine gömülü bloklardan elde edilen DNA izolasyonunu takiben, 28 adet kanser ilişkili gene ait genetik değişiklikler yeni nesil dizileme analizi (NGS) ile tayin edildi. Dokulardaki TNF- α ekspresyonu immünhistokimyasal çalışmalar ile tayin edildi. İnflamasyon skorlaması, her vaka için hemotoksilen-eosin kesitlerde yapıldı. Toplam 25 adet OSHK vakasının 18’inde (%72) bir veya birden fazla mutasyon izlenmiştir. En sık mutasyonu görülen gen, P53 geni olarak saptanmıştır. Tüm vakalardan 3 tanesi (%12) üç ve daha fazla gen mutasyonu göstermiştir. TNF- α ekspresyonu mutasyon saptanan tümörlerde daha fazla izlenmiştir. Yeni nesil dizileme analizi (NGS) ile ortaya çıkan mutasyon spektrumu, OSHK'deki genetik değişikliklerin ne kadar çeşitli ve karmaşık olduğunu göstermektedir. Çalışma sonuçları genetik değişiklikler ile inflamasyon ve TNF-α ekspresyonu arasında ilişki olduğunu düşündürmektedir. OSHK’nin tanı ve tedavisinde histopatolojik incelemenin yanı sıra, genetik alterasyonları da ortaya koymak hedefe yönelik tedavide yeni alternatifler sunacaktır.

Anahtar Kelimeler

oral cancer, yeni nesil dizileme, TNF-a

Kaynakça

• 1. Jayaprakash C, Varghese VK, Jayaram P, Chakrabarty S, Kudva A, Ray S, et al. Relevance and actionable mutational spectrum in oral squamous cell carcinoma. J Oral Pathol Med. 2019;(July):1–8.
• 2. Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer. Oral Oncol [Internet]. 2009;45(4–5):309–16. Available from: http://dx.doi.org/10.1016/j.oraloncology.2008.06.002
• 3. Hanahan D, Weinberg RA. Hallmarks of cancer: The next generation. Cell [Internet]. 2011;144(5):646–74. Available from: http://dx.doi.org/10.1016/j.cell.2011.02.013
• 4. Goertzen C, Mahdi H, Laliberte C, Meirson T, Eymael D, Gil-Henn H, et al. Oral inflammation promotes oral squamous cell carcinoma invasion. Oncotarget. 2018;9(49):29047–63.
• 5. Krishnan R, Thayalan DK, Padmanaban R, Ramadas R, Annasamy RK, Anandan N. Association of serum and salivary tumor necrosis factor-α with histological grading in oral cancer and its role in differentiating premalignant and malignant oral disease. Asian Pacific J Cancer Prev. 2014;15(17):7141–8.
• 6. Dantas TS, de Barros Silva PG, Verde MEQL, Ribeiro Júnior A de L, Cunha M do PSS, Mota MRL, et al. Role of inflammatory markers in prognosis of oral squamous cell carcinoma. Asian Pacific J Cancer Prev. 2019;20(12):3635–42.
• 7. Deepthi G, Nandan SRK, Kulkarni PG. Salivary tumour necrosis factor-α as a biomarker in oral leukoplakia and oral squamous cell carcinoma. Asian Pacific J Cancer Prev. 2019;20(7):2087–93.
• 8. Nakano Y, Kobayashi W, Sugai S, Kimura H, Yagihashi S. Expression of tumor necrosis factor-α and interleukin-6 in oral squamous cell carcinoma. Japanese J Cancer Res. 1999;90(8):858–66.
• 9. Scheff NN, Ye Y, Bhattacharya A, Macrae J, Hickman DH, Atul K, et al. inflammation. Vol. 158. 2018. 2396–2409 p.
• 10. Yangın S, Tanman Zıplar Ü, Cansaran-Duman D. Pharmaceutical Applications Based on Next Generation Sequencing Technology in Oncologic Drug Development. Turkish Bull Hyg Exp Biol. 2019;76(4):473–86.
• 11. Sharma V, Nandan A, Sharma AK, Singh H, Bharadwaj M, Sinha DN, et al. Signature of genetic associations in oral cancer. Tumor Biol. 2017;39(10):1–9.
• 12. Jessri M, Farah CS. Next generation sequencing and its application in deciphering head and neck cancer. Oral Oncol [Internet]. 2014;50(4):247–53. Available from: http://dx.doi.org/10.1016/j.oraloncology.2013.12.017
• 13. Nakagaki T, Tamura M, Kobashi K, Koyama R, Fukushima H, Ohashi T, et al. Profiling cancer-related gene mutations in oral squamous cell carcinoma from Japanese patients by targeted amplicon sequencing. Oncotarget. 2017;8(35):59113–22.
• 14. Hagemann IS, Devarakonda S, Lockwood CM, Spencer DH, Guebert K, Bredemeyer AJ, et al. Clinical next-generation sequencing in patients with non-small cell lung cancer. Cancer. 2015;121(4):631–9.
• 15. Thompson LDR. World health organization classification of tumours: Pathology and genetics of head and neck tumours. Ear, Nose Throat J. 2006;85(2):74. 16. Sawazaki-Calone I, Rangel ALCA, Bueno AG, Morais CF, Nagai HM, Kunz RP, et al. The prognostic value of histopathological grading systems in oral squamous cell carcinomas. Oral Dis. 2015;21(6):755–61.
• 17. Loyo M, Li RJ, Bettegowda C, Pickering CR, Frederick MJ, Myers JN, et al. Lessons learned from next-generation sequencing in head and neck cancer. Head Neck. 2013;35(3):454–63.
• 18. Cara B. Gonzalesa, b,*, Jorge J. De La Chapab, Pothana Saikumarc, Prajjal K. Singhac, Nicholas F. Dybdal-Hargreavesd, Jeffery Chaveze, Aaron M. Horningf, Jamie Parrac and NBK. Co-targeting ALK and EGFR parallel signaling in oral squamous cell carcinoma. Oral Oncol 2016 August ; 59 12–19 doi101016/j.oraloncology201605007
• 19. Xu J, Lu W, Zhang S, Zhu C, Ren T, Zhu T, et al. Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma. Cancer Biol Ther. 2014;15(5):612–22.