Kedi skuamöz hücreli karsinomlarının transkripsiyonel ekspresyon profili

Yıl 2025, Cilt: 10 Sayı: 6, 861 - 868, 30.11.2025

Mahmut Sözmen , Yonca Betil Kabak , Sinem İnal , Fatma Betül Akça

https://doi.org/10.35229/jaes.1775035

Öz

Skuamöz hücreli karsinom (SCC), kedilerde en sık görülen malign kutanöz ve oral tümörlerdir. Kedi skuamöz hücreli karsinomlarında (FSCC) genomik değişiklikler kedilerde karakterize edilmemiştir. Bu çalışmanın amacı, mikroarray analizi ile doğal olarak ortaya çıkan FSCC vakalarında ekspresyonu önemli ölçüde değişen genleri belirlemektir. İki adet formalinle fikse edilmiş parafin gömülü (FFPE) FSCC vakası, GeneChipTM Feline Gene 1.0 ST Array ile analiz edildi. Mikroarray analizi, FSCC vakalarının 35.034 gen düzeyinde prob setinden 148 farklı şekilde ifade edilen gen barındırdığını ortaya koydu. Bu değişikliklerden 136 gen yukarı doğru düzenlenirken, diğer 12 gen aşağı doğru düzenlendi. Mikroarray analizi sonuçları, yukarı regüle edilen genlerin çoğunun onkojenik olduğunu ortaya koydu. FSCC gelişimine yol açan moleküler olaylarda rol oynayabilecek, ekspresyon seviyeleri önemli ölçüde yükselmiş genler (NCK1, PDE7A, UBAP2L, PPP2R5A ve CDK1) seçildi ve bölüm arşivlerinden alınan toplam 26 FFPE FSCC vakasında gerçek zamanlı PCR ile daha ayrıntılı olarak incelendi. Buna göre, bu 26 FFPE FSCC vakasında, NCK1, PDE7A, UBAP2L, PPP2R5A ve CDK1 genlerinin kat değişiklikleri sırasıyla 6,96, 2,58, 5,24, 55,13 ve 3,5 idi. Mikroarray analizi ve gerçek zamanlı PCR sonuçları, yukarı regüle edilen genlerin hücre döngüsünde rol oynadığını ve proliferasyonun da FSCC'nin gelişimine katkıda bulunabileceğini göstermektedir.

Anahtar Kelimeler

gen ifadesi , kedi , mikroarray , skuamöz hücreli karsinom.

Proje Numarası

TUBITAK-TOVAG; Project No.: 119O708

Transcriptional expression profiling of feline squamous cell carcinomas

Öz

Squamous cell carcinoma (SCC) is the most common malignant cutaneous and oral tumors occurring in cats. Genomic alterations in feline squamous cell carcinomas (FSCC) have not been characterized in cats. The objective of the present study was to determine genes of which expression was significantly altered in naturally occurring FSCC cases by microarray analysis. Two formalin fixed paraffin embedded (FFPE) FSCC cases were analyzed with GeneChipTM Feline Gene 1.0 ST Array. Microarray analysis revealed that FSCC cases harbored 148 differentially expressed genes out of 35,034 gene-level probe sets. Of these changes, 136 genes were up-regulated while the other 12 genes were down-regulated. Microarray analysis results revealed that most of the upregulated genes were oncogenic. Expression levels significantly elevated genes (NCK1, PDE7A, UBAP2L, PPP2R5A, and CDK1) which may have a role in the molecular events leading to the development of FSCC were selected and further examined by real-time PCR in a total of 26 FFPE FSCC cases retrieved from departmental archives. Accordingly, in those 26 FFPE FSCC cases, the fold changes for the NCK1, PDE7A, UBAP2L, PPP2R5A, and CDK1 genes were 6.96, 2.58, 5.24, 55.13, 3.5, respectively. Microarray analysis and real-time PCR results indicate that upregulated genes are involved in the cell cycle and proliferation may also contribute to the development of FSCC.

Anahtar Kelimeler

cat , gene expression , microarray , squamous cell carcinoma

Destekleyen Kurum

Funding: This work was financially supported by the Turkish Scientific Research Council (TUBITAK-TOVAG; Project No.: 119O708), Ankara, Turkey.

Proje Numarası

TUBITAK-TOVAG; Project No.: 119O708

Kaynakça

• Andersson, E.M., Paoli, J., & Wastensson, G. (2011). Incidence of cutaneous squamous cell carcinoma in coastal and inland areas of Western Sweden. Cancer Epidemiol, 35, e69-e74. DOI: 10.1016/j.canep.2011.05.006
• Brewster, D.H., Bhatti, L.A., Inglis, J.H., Nairn, E.R., & Doherty, V.R. (2007). Recent trends in incidence of nonmelanoma skin cancers in the East of Scotland, 1992-2003. Br. J. Dermatol., 156, 1295-1300. DOI: 10.1111/j.1365-2133.2007.07892.x
• Chai, R., Yu, X., Tu, S., & Zheng, B. (2016). Depletion of UBA protein 2-like protein inhibits growth and induces apoptosis of human colorectal carcinoma cells. Tumour Biol, 37, 13225-13235. DOI: 10.1007/s13277-016- 5159-y
• Cheong, J.Y., Kim, Y.B., Woo, J.H., Kim, D.K., Yeo, M., Yang, S., Yang, K., Sun, S.K., Wang, H.J., Kim, B.W., Park, J., & Cho, S.W. (2016). Identification of NUCKS1 as a putative oncogene and immunodiagnostic marker of hepatocellular carcinoma. Gene, 584, 47-53. DOI: 10.1016/j.gene.2016.03.006
• Drosos, Y., Kouloukoussa, M., Østvold, A.C., Grundt, K., Gouta, N., Vlachodimitropoulos, D., Havaki, S., Kollia, P., Kittas, C., Marinos, E., & Aleporou- Marinou, V. (2009). NUCKS overexpression in breast cancer. Cancer Cell Int, 9, 19. DOI: 10.1186/1475- 2867-9-19
• Goldschmidt, M.H., Dunstan, R.W., Stannard, A.A., von Tscharner, C., Walder, E.J., & Yager, J.A. (1998). Histological classification of epithelial and melanocytic tumors of the skin of domestic animals, 2nd ed, Armed Forces Institute of Pathology, Washington.
• Goldschmidt, M.H., & Shofer, F.S. (1992). Squamous cell carcinoma. In: Goldschmidt, M.H., & Shofer, F.S.(Eds) Skin tumors of the dog and the cat. 1st ed, Pergamon Press: New York; 37-50p.
• Goto, M., Kadoshima-Yamaoka, K., Murakawa, M., Yoshioka, R., Tanaka, Y., Inoue, H., Murafuji, H., Kanki, S., Hayashi, Y., Nagahira, K., Ogata, A., Nakatsuka, T., & Fukuda, Y. (2010). Phosphodiesterase 7A inhibitor ASB16165 impairs proliferation of keratinocytes in vitro and in vivo. Eur J Pharmacol, 633, 93-97. DOI: 10.1016/j.ejphar.2010.01.024
• Gross, T.L., Ihrke, P.J., Walder, E.J., & Affolter, V.K. (2005). Squamous cell carcinoma. In: Gross, T.L., Ihrke, P.J., Walder, E.J., & Affolter, V.K. (EDs), Skin diseases of the dog and cat. 2nd ed, Blackwell: Iowa, 581-589p.
• Gu, L., Xia, B., Zhong, L., Ma, Y., Liu, L., Yang, L., & Lou G. (2014). NUCKS1 overexpression is a novel biomarker for recurrence-free survival in cervical squamous cell carcinoma. Tumour Biol, 35, 7831-7836. https://doi.org/10.1007/s13277-014-2035-5
• He, J., Chen, Y., Cai, L., Li, Z., & Guo, X. (2018). UBAP2L silencing inhibits cell proliferation and G2/M phase transition in breast cancer. Breast Cancer, 25, 224-232. DOI: 10.1007/s12282-017-0820-x
• Huang, Y.K., Kang, W.M., Ma, Z.Q., Liu, Y.Q., Zhou, L., & Yu, J.C. (2018). NUCKS1 promotes gastric cancer cell aggressiveness by upregulating IGF-1R and subsequently activating the PI3K/Akt/mTOR signaling pathway. Carcinogenesis, 40, 370-379. DOI: 10.1093/carcin/bgy142
• Kabak, Y.B., Sözmen, M., Devrim, A.K., Sudağıdan, M., Yıldırım, F., Güvenç, T., Yarım, M., Gülbahar, M.Y., Ahmed, I., Karaca, E., & İnal, S. (2020). Expression levels of angiogenic growth factors in the feline squamous cell carcinoma. Acta Vet Hungarica, 68, 37-48. DOI: 10.1556/004.2020.00005
• Katalinic, A., Kunze, U., & Schafer, T. (2003). Epidemiology of cutaneous melanoma and non-melanoma skin cancer in Schleswig Holstein, Germany: incidence, clinical subtypes, tumour stages and localization (epidemiology of skin cancer). Br. J. Dermatol., 149, 1200-1206. DOI: 10.1111/j.1365-2133.2003.05554.x
• Lambert, S.R., Mladkova, N., Gulati, A., Hamoudi, R., Purdie, K., Cerio, R., Leigh, I., Proby, C., & Harwood, C.A. (2014). Key differences identified between actinic keratosis and cutaneous squamous cell carcinoma by transcriptome profiling. Br. J. Cancer, 110, 520-529. DOI: 10.1038/bjc.2013.760
• Li, H., Li, B., & Larose, L. (2017). IRE1 alpha links Nck1 deficiency to attenuated PTP1B expression in HepG2 cells. Cell Signal, 36, 79-90. DOI: 10.1016/j.cellsig.2017.04.017
• Maeda, M., Hasegawa, H., Sugiyama, M., Hyodo, T., Ito, S., Chen, D., Asano, E., Masuda, A., Hasegawa, Y., Hamaguchi, M., & Senga, T. (2016). Arginine methylation of ubiquitin-associated protein 2-like is required for the accurate distribution of chromosomes. FASEB J, 30, 312-323. DOI: 10.1096/fj.14-268987
• Mao, Z., Liu, C., Lin, X., Sun, B., & Su, C. (2018). PPP2R5A: A multirole protein phosphatase subunit in regulating cancer development. Cancer Lett., 414, 222-229. DOI: 10.1016/j.canlet.2017.11.024
• Marko D, Romanakis K, Zankl H, Fürstenberger G, Steinbauer B, & Eisenbrand G. (1998). Induction of apoptosis by an inhibitor of cAMP-specific PDE in malignant murine carcinoma cells overexpressing PDE activity in comparison to their non-malignant counterparts. Cell Biochem. Biophys, 28, 75-101. DOI: 10.1007/BF02737806
• Miller, M.A., Nelson, S.L., Turk, J.R., Pace, L.W., Brown, T.P., Shaw, D.P., Fischer, J.R., & Gosser, H.S. (1991). Cutaneous neoplasia in 340 cats. Vet Pathol, 28, 389-395. DOI: 10.1177/030098589102800506
• Miyamoto, Y., Torii, T., Kawahara, K., Tanoue, A., & Yamauchi, J. (2016). Dock8 interacts with Nck1 in mediating Schwann cell precursor migration. Biochem Biophy Rep, 6, 113-123. DOI: 10.1016/j.bbrep.2016.03.013
• Muzic, J.G., Schmitt, A.R., Wright, A.C., Alniemi, D.T., Zubair, A.S., Olazagasti Lourido J.M., Sosa Seda, I.M., Weaver, A.L., & Baum, C.L. (2017). Incidence and trends of basal cell carcinoma and cutaneous squamous cell carcinoma: a population-based study in Olmsted County, Minnesota, 2000 to 2010. Mayo Clin Proc, 92, 890-898. DOI: 10.1016/j.mayocp.2017.02.015
• Nindl, I., Dang, C., Forschner, T., Kuban, R.J., Meyer, T., Sterry, W., & Stockfleth, E. (2006). Identification of differentially expressed genes in cutaneous squamous cell carcinoma by microarray expression profiling. Mol Cancer, 5, 30. DOI: 10.1186/1476-4598-5-30
• Okamoto, K., Li, H., Jensen, M.R., Zhang, T., Taya, Y., Thorgeirsson, S.S., & Prives, C. (2002). Cyclin G recruits PP2A to dephosphorylate Mdm2. Mol Cell, 9, 761-771. DOI: 10.1016/s1097-2765(02)00504-x
• Ostvold, A.C., Norum, J.H., Mathiesen, S., Wanvik, B., Sefland, I., & Grundt, K. (2001). Molecular cloning of a mammalian nuclear phosphoprotein NUCKS, which serves as a substrate for Cdk1 in vivo. Eur J Biochem, 268, 2430-2440. DOI: /10.1046/j.1432- 1327.2001.02120.x
• Pontius, J.U., Mullikin, J.C., Smith, D.R., Agencourt Sequencing Team, Lindblad-Toh, K., Gnerre, S., Clamp, M., Chang, J., Stephens, R., Neelam, B., Volfovsky, N., Schäffer, A.A., Agarwala, R., Narfström, K., Murphy, W.J., Giger, U., Roca, A.L., Antunes, A., Menotti-Raymond, M., Yuhki, N., Pecon-Slattery, J., Johnson, W.E., Bourque, G., Tesler, G., NISC Comparative Sequencing Program, & O’Brien, S.J. (2007). Initial sequence and comparative analysis of the cat genome. Genome Res, 17, 1675-1689. DOI: 10.1101/gr.6380007
• Schaner, M.E., Ross, D.T., Ciaravino, G., Sørlie, T., Troyanskaya, O., Diehn, M., Wang, Y.C., Duran, G.E, Sikic, T.L., Caldeira, S., Skomedal, H., Tu, I.P., Hernandez-Boussard, T., Johnson, S.W., O’Dwyer, P.J., Fero, M.J., Kristensen, G.B., Børresen-Dale, A.L., Hastie, T., Tibshirani, R., van de Rijn, M., Teng, N.N., Longacre, T.A., Botstein, D., Brown, P.O., & Sikic, B.I. (2003). Gene expression patterns in ovarian carcinomas. Mol. Biol. Cell, 14, 4376-4386. DOI: 10.1091/mbc.e03-05-0279
• Supsavhad, W., Dirksen, W.P., Martin, C.K., & Rosol, T.J. (2016). Animal models of head and neck squamous cell carcinoma. The Veterinary Journal, 210, 7-16. DOI: 10.1016/j.tvjl.2015.11.006
• Thomas, R., Valli, V.E., Ellis, P., Bell, J., Karlsson, E.K., Cullen, J., Lindblad-Toh, K., Langford, C.F., & Breen, M. (2009). Microarray-based cytogenetic profiling reveals recurrent and subtype-associated genomic copy number aberrations in feline sarcomas. Chromosome Res, 17, 987-1000. DOI: 10.1007/s10577-009-9096-0
• Walaas, S.I., Ostvold, A.C., & Laland, S.G. (1989). Phosphorylation of P1, a high mobility group-like protein, catalyzed by casein kinase II, protein kinase C, cyclic AMP-dependent protein kinase and calcium/ calmodulin-dependent protein kinase II. FEBS Lett, 258, 106-108. DOI: 10.1016/0014-5793(89)81626-6
• Wei, W., Chen, Y., Xu, J., Zhou, Y., Bai, X., Yang, M., & Zhu, J. (2018). Identification of biomarker for cutaneous squamous cell carcinoma using microarray data analysis. J. Cancer, 9, 400-406. DOI: 10.7150/jca.21381
• Wilcock, B.P. (2006). Eye and ear. In: Maxie, MG. (Ed). Jubb, Kennedy, and Palmer’s pathology of domestic animals. Saunders: Philadelphia, 534-536p.
• Wisniewski, J.R., & Schwanbeck, R. (2000). High mobility group I/Y: multifunctional chromosomal proteins causally involved in tumor progression and malignant transformation. Int. J. Mol. Med., 6, 409-419. DOI: 10.3892/ijmm.6.4.409
• Xia, P., Huang, M., Zhang, Y., Xiong, X., Yan, M., Xiong, X., Yu, W., & Song, E. (2019). NCK1 promotes the angiogenesis of cervical squamous carcinoma via Rac1/PAK1/MMP2 signal pathway. Gynecol Oncol, 152, 387-395. DOI: 10.1016/j.ygyno.2018.11.013
• Yang, Y., Kitagaki, J., Wang, H., Hou, D.X., & Perantoni, A.O. (2009). Targeting the ubiquitin-proteasome system for cancer therapy. Cancer Sci, 100, 24-28. DOI: 10.1111/j.1349-7006.2008.01013.x
• Ye, T., Xu, J., Du, L., Mo, W., Liang, Y., & Xia, J. (2017). Downregulation of UBAP2L inhibits the epithelial- mesenchymal transition via SNAIL1 regulation in hepatocellular carcinoma cells. Cell Physiol. Biochem., 41, 1584-1595. DOI: 10.1159/000470824
• Zhang, F., Lu, Y.X., Chen, Q., Zou, H.M., Zhang, J.M., Hu, Y.H., Li, X.M., Zhang, W.J., Zhang, W., Lin, C., & Li, X.N. (2017). Identification of NCK1 as a novel downstream effector of STAT3 in colorectal cancer metastasis and angiogenesis. Cell Signal, 36, 67-78. DOI: 10.1016/j.cellsig.2017.04.020
• Zhang, L., Murray, F., Zahno, A., Kanter, J.R., Chou, D., Suda, R., Fenlon, M., Rassenti, L., Cottam, H., Kipps, T.J., & Insel, P.A. (2008). Cyclic nucleotide phosphodiesterase profiling reveals increased expression of phosphodiesterase 7B in chronic lymphocytic leukemia. Proc Natl Acad Sci USA, 105, 19532-19537. DOI: 10.1073/pnas.0806152105
• Zhang, L., Qin, H., Wu, Z., Chen, W., & Zhang, G. (2018). Pathogenic genes related to the progression of actinic keratosis to cutaneous squamous cell carcinoma. Int J Dermatol, 57,120.