RETİNOİK ASİT TESTİS EMBRİYONEL KARSİNOMUNDAN NÖRONA BİR GEÇİŞ YOLU AÇAR MI?

Year 2025, Volume: 6 Issue: 3, 212 - 219, 25.11.2025

Hilal Nakkaş , Şeyma Kipel

Abstract

Giriş: Testis embriyonal karsinomu, sperm oluşumuna yol açan germ hücrelerini etkileyen testis kanseri türlerinden birisidir. Bu kanser hücreleri kendini yenileyebilir, farklılaşabilir ve kötü huylu tümörlerin büyümesini teşvik eder. A vitamininin (retinol) aktif bir yan ürünü olan retinoik asit (RA), hem embriyonik gelişim hem de hücre bölünmesi, çoğalması ve ölümü gibi temel biyolojik işlevler için önemlidir. Özellikle kanser ve kök hücrelerde, in vitro farklılaşma indükleyicisi olarak sıklıkla kullanılır. S100B proteini, iltihaplanma, hücre büyümesi, hücre farklılaşması, hücre iskeleti dinamikleri ve hücre hareketi gibi olaylarda önemli rol oynar. NF-κB (nükleer faktör kappa B), hücre içinde önemli bir transkripsiyon faktörü olan bir protein kompleksidir. Sürekli aktif olan NF-κB, bazı kanserlerde hücre çoğalmasını artırır ve bağışıklık sisteminin tümöre verdiği yanıtı engeller. Çalışmanın amacı, RA uyarımı sonucu testis embriyonal karsinom hücrelerinin morfolojileri ile S100B ve NF-κB ekspresyonlarının nasıl değiştiğini göstermektir.

Yöntem: Testis embriyonal karsinom hücrelerine (CRL-2073) belirlenen dozda (10 μM) RA uygulandıktan sonra, morfolojik inceleme için kristal viyole ve luxol fast blue boyamaları yapıldı. Daha sonra, immünohistokimyasal teknik kullanılarak, testis embriyonal karsinom hücrelerinde S100B ve NF-κB'nin hücresel ekspresyonu ve yeri incelendi.

Bulgular: RA ile farklılaşan CRL-2073 hücrelerini kristal viyole ile boyadığımızda, hücre çekirdeğinde ve sitoplazmada farklılaşmanın görsel bir göstergesi olan morfolojik farklılıklar gözlemledik. RA ile nöronal farklılaşmaya başlayan CRL-2073 hücrelerinde luxol fast blue boyaması gözlendi. S100B proteini embriyonal karsinom hücrelerinde ifade edildi ve hücre farklılaşmasıyla ilişkili bulundu. NF-κB bu hücrelerde proliferasyon ve pluripotensiyi korumada aktiftir; aktivitesi farklılaşmayla azaldı.

Sonuç: CRL-2073 hücrelerinin RA ile farklılaşmaya başladığını histolojik olarak göstermek önemlidir. Ayrıca, hem tümör biyolojisi hem de farklılaşma süreçleri için biyobelirteç olan S100B ve NF-κB proteinlerinin ifade düzeylerinin RA öncesi ve sonrasında immünboyama ile belirlenmesi potansiyel bir tedavi hedefidir. Bu bilgi kanser biyolojisini anlamada önemli bir adımdır.
Anahtar kelimeler: Testis embriyonal karsinom, retinoik asit, farklılaşma, S100B, NF-κB.

Keywords

Testis embriyonal karsinomu , retinoik asit , farklılaşma , S100B , NF-κB.

DOES RETINOIC ACID PAVE A WAY FROM TESTICULAR EMBRYONAL CARCINOMA TO NEURON?

Abstract

Introduction: One kind of testicular cancer which affects germ cells that eventually give rise to sperm is called testicular embryonal carcinoma. These cancer cells can self-renew and differentiate, and they encourage the growth of malignancies. An active byproduct of vitamin A (retinol), retinoic acid (RA), is crucial for both embryonic development and fundamental biological functions such cell division, proliferation, and death. It is frequently utilized as a differentiation inducer in vitro, particularly on cancer and stem cells. S100B protein plays an important role in events such as inflammation, cell growth, cell differentiation, cytoskeleton dynamics and cell movement. NF-κB (nuclear factor kappa B) is a protein complex that is an important transcription factor within the cell. Constantly active NF-κB increases cell proliferation in some cancers and inhibits the immune system's response to the tumor. The aim of the study is to demonstrate how the morphology and S100B and NF-κB expressions of testicular embryonal carcinoma cells change as a result of RA stimulation.

Method: After RA was administered to the testicular embryonal carcinoma cells (CRL-2073) at the determined dose (10 μM), crystal violet and luxol fast blue stainings were performed for morphological examination. Then, using immunohistochemical technique, cellular expression and location of S100B and NF-κB in testicular embryonal carcinoma cells were examined.

Results: When we stained CRL-2073 cells differentiated with RA with crystal violet, we observed morphological differences in the cell nucleus and cytoplasm, which is a visual indicator of differentiation. Luxol fast blue staining was observed in CRL-2073 cells that began neuronal differentiation with RA. The S100B protein was expressed in embryonal carcinoma cells and was associated with cell differentiation. NF-κB was active in maintaining proliferation and pluripotency in these cells; its activity decreased with differentiation.

Conclusion: It is important to show histologically that CRL-2073 cells begin to differentiate with RA. In addition, determining the expression levels of S100B and NF-κB proteins, which are biomarkers for both tumor biology and differentiation processes, by immunostaining before and after RA is a potential treatment target. This information is an important step in understanding cancer biology.

Keywords

Testis embryonal carcinoma , retinoic acid , differantiation , S100B , NF-κB.

Ethical Statement

Çalışmamız hücre hattı üzerinde yapıldığı için etik kurul onay belgesi yoktur. Bilgilerinize sunarım. Saygılarımla.

References

• Reuter V. Origins and molecular biology of testicular germ cell tumors. Modern Pathology 2005; 18(S2), S51–S60.
• Baird D, Meyers G, Hu J. Testicular cancer: Diagnosis and treatment. American Family Physician 2018; 97(4), 261-8.
• Yamanaka S. Pluripotent stem cell-Based cell therapy-Promise and Challenges. Cell Stem Cell 2020; 523-31.
• Teshima S, Shimosato Y, Hirohashi S et al. Four new human germ cell lines. Lab Investig 1988; 59:328–36.
• Damjanov I, Horvat B, Gibas Z. Retinoic acid-induced differentiation of the developmentally pluripotent human germ cell tumor-derived cell line, NCCIT. Lab Investig 1993; 68:220–32.
• Soltanian S, Sheikhbahaei M, Ziasistani M. Phytol down regulates expression of some cancer stem cell markers and decreases side population proportion in human embryonic carcinoma NCCIT cells. Nutrion and Cancer 2021; 1520-33.
• Janesick A, Cherie W, Blumberg B. Retinoic acid signaling and neuronal differentiation. Cell Mol Life 2015; 72 (8): 1559-76.
• Chen H, Xu C, Jin Q et al. S100 protein family in human cancer. American Journal of Cancer Research 2014, 4(2), 89-115.
• Gonzalez L, Garrie K, Turner M. Role of S100 proteins in health and disease. Biochmica et Biophysica Acta (BBA)-Molecular Cell Research 2020;118677.
• Tsutsui Y, Nogami T, Kashiwai A et al. Induction of S100b (beta beta) protein in human teratocarcinoma cells. Cell Differ 1987; 21(2):137-45.
• Wang S, Rosegren L, Franlund M et al. Bcl-2 expression regulates cell sensivity to S100beta-mediated apoptosis. Brain Res Mol Brain Res 1999; 70(1);167-76.
• Sung B, Do H, Park S et al. Regulation of OCT4 gene expression by liver receptor homolog-1 in human embryonic carcinoma cells. Biochemical and Biophysical Research Communications 2012; 427(2), 315–20.
• Coyle D, Li J, Baccei M. Regional differentiation of retinoic acid- induced human pluripotent embryonic carcinoma stem cell neurons. Plos One 2011; 20(6);16174.
• Imran M, Park J, Lim I. Stress-induced NF-KB activation differentiates promyelocytic leukemia cells to macrophages in response to all-trans-retinoic acid. Cell Signaling 2015; 27(3):694-706.
• Gasimli L, Stansfield H, Nairn A et al. Structural remodeling of proteoglycans upon retinoic acid-induced differentiation of NCCIT cells. Glycoconj 2013; 30(5):497-510.
• Park S, Do H, Ha W et al. Differential Expression of ETS Family Transcription Factors in NCCIT Human Embryonic Carcinoma Cells upon Retinoic Acid-Induced Differentiation. Biological and Pharmaceutical Bulletin 2014; 37(4), 659–65.
• Niu C, Li M, Ni Y et al. Effect of all-trans retinoci acid on the proliferation and differentiation of brain tumor stem cells. Journal of Experimental &Clinic Cancer Reserach 2010; 29(1):113.
• Joo J, Kim J, Lee Y et al. Involment of NK-kappaB in the regulation of S100A6 gene expression in human hepatoblastoma cell line HepG2. Biochem Biophys Res. Commun 2003; 307(2): 274-80.
• Spinella M, Freemantle S, Sekula D et al. Retinoic acid promotes ubiquitination and proteolysis of cyclin D1 during induced tumor cell differentiatşon. Cell Biology Metabolism 1999;22013-8.
• Baldassarre G, Bruni A, Sandemenico C et al. Retinoic acid induces neuronal differentiation of embryonal carcinoma cells by reducing proteasome dependent proteolysis of the cyclin dependent inhibitör p27. Cell Growth Differ 2000; 11(10): 517-26.
• JH S, Bours N, JA M et al. Retinoic acid induction of majör histocompatibility complex class I genes in NTera-2 embriyonal carcinoma cell involves induction of NF-kappa B (p50-p6 5) and retinoic acid receptor beta retinoid X receptor beta heterodimers. Molecular and Cellular Biology 1993; 13(10): 6157-69.
• Park S, Do H, Ha W et al. Transcriptional regulation of OCT4 by the ETS transcription factor ESE-1 in NCCIT human embriyonic carcinoma cells. Biochemical and Biophysical Reserach Communications 2014; 984-90.