EFFECTS OF ADIPOSE-DERIVED STEM CELL - CONDITIONED MEDIUM ON CoCl₂ - ADAPTED ORAL SQUAMOUS CELL CARCINOMA CELLS

Öz

Objective: The progression of oral squamous cell carcinoma (OSCC) is strongly influenced by hypoxia and signals derived from the tumor microenvironment. This study aimed to investigate the effects of adipose-derived stem cell-conditioned medium (ASC-CM) on CoCl₂-induced hypoxic UPCI-SCC-131 cells. Methods: Hypoxia induction was confirmed by significant upregulation of HIF1A and CA9 gene expression and increased Hif1α protein levels. The effects of ASC-CM on cell viability were assessed, and the expression of genes associated with cell survival and stress responses was analyzed by RT-qPCR. Intracellular reactive oxygen species (ROS) production, migratory capacity, spheroid formation, and sensitivity to cisplatin were also evaluated. Results: ASC-CM slightly reduced cell viability while markedly altering cellular behavior. RT-qPCR analysis revealed downregulation of the cell survival-related genes APAF1, CASP2, CASP9, and BAX, accompanied by increased BCL2 expression. In addition, the expression levels of HMOX1, CCL2, HIF1A, and CA9 were significantly increased, suggesting activation of oxidative stress- and survival-associated responses. Functionally, ASC-CM enhanced intracellular ROS production, increased migratory and spheroid formation capacity, and reduced sensitivity to cisplatin. Conclusion: These findings suggest that ASC-derived conditioned medium modulates oxidative stress- and cell survival-associated responses and influences several cellular behaviors associated with tumor progression in hypoxia-mimicking OSCC cells. The results further indicate that factors derived from the tumor microenvironment may contribute to the development of a more aggressive phenotype in hypoxic OSCC cells.

Anahtar Kelimeler

• oral squamous cell carcinoma
• adipose-derived stem cells
• conditioned medium
• hypoxia
• cisplatin resistance
• ROS

Etik Beyan

This study was conducted in accordance with the principles of research and publication ethics. No human participants or animals were involved in this study; therefore, ethical committee approval and informed consent were not required. The authors declare no conflict of interest.

Adipöz Kaynaklı Kök Hücre Koşullandırılmış Ortamının CoCl₂ ile Adapte Edilmiş Oral Skuamöz Hücreli Karsinom Hücrelerinde Agresif Hücresel Özellikler Üzerindeki Etkileri

Öz

Amaç: Oral skuamöz hücreli karsinomun (OSHK) ilerlemesi, hipoksi ve tümör mikroçevresinden kaynaklanan sinyallerden önemli ölçüde etkilenmektedir. Bu çalışmada, adipöz kaynaklı kök hücre koşullandırılmış ortamının (ASC-CM), CoCl₂ ile hipoksi benzeri koşullara adapte edilmiş UPCI-SCC-131 hücreleri üzerindeki etkilerinin araştırılması amaçlandı. Yöntem: Hipoksi indüksiyonu, HIF1A ve CA9 gen ekspresyonlarındaki artış ile Hif1α protein düzeylerinin yükselmesiyle doğrulandı. ASC-CM uygulamasının hücre canlılığı üzerindeki etkisi değerlendirildi. Hücre sağkalımı ve stres yanıtı ile ilişkili genlerin ekspresyonları RT-qPCR yöntemiyle analiz edildi. Ayrıca hücre içi reaktif oksijen türleri (ROS) üretimi, migrasyon kapasitesi, sferoid oluşumu ve sisplatine duyarlılık incelendi. Bulgular: ASC-CM, hücre canlılığını hafif düzeyde azaltırken hücresel davranışları belirgin şekilde değiştirdi. RT-qPCR analizleri, hücre sağkalımı ile ilişkili gen ekspresyonlarında değişiklikler olduğunu gösterdi; APAF1, CASP2, CASP9 ve BAX ekspresyonları azalırken BCL2 ekspresyonu arttı. HMOX1, CCL2, HIF1A ve CA9 ekspresyon düzeylerinde anlamlı artışlar saptandı. ASC-CM uygulaması hücre içi ROS üretimini artırdı, migrasyon ve sferoid oluşturma kapasitesini güçlendirdi ve hücrelerin sisplatine duyarlılığını azalttı. Sonuç: Elde edilen bulgular, adipöz kaynaklı kök hücre koşullandırılmış ortamının hipoksi benzeri koşullara adapte edilmiş OSHK hücrelerinde oksidatif stres ve hücre sağkalımı ile ilişkili yanıtları modüle ettiğini ve tümör progresyonu ile ilişkili çeşitli hücresel özellikleri etkilediğini göstermektedir. Bu sonuçlar, tümör mikroçevresinden kaynaklanan faktörlerin hipoksik OSHK hücrelerinde daha agresif bir fenotipin gelişimine katkıda bulunabileceğini düşündürmektedir.

Anahtar Kelimeler

• oral skuamöz hücreli karsinom
• adipöz kaynaklı kök hücreler
• koşullandırılmış ortam
• hipoksi
• sisplatin direnci
• reaktif oksijen türleri (ROS)
• migrasyon

Etik Beyan

Bu çalışma araştırma ve yayın etiği ilkelerine uygun olarak gerçekleştirilmiştir. Çalışmada insan katılımcılar veya deney hayvanları kullanılmamış olup, yalnızca hücre kültürü modelleri kullanılmıştır. Bu nedenle etik kurul onayı ve bilgilendirilmiş onam gerekmemektedir. Yazarlar herhangi bir çıkar çatışması bulunmadığını beyan eder.

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