The role of Odoroside A in immune transformation in RAW264.7 cells

Öz

Purpose: Macrophages are one of the most important elements of the immune system and play dual role in inflammation. They regulate both the formation of the immune response in the environment and the suppression. This signaling in the cancer microenvironment affects the progression of cancer. This cancer-promoting or cancer-eliminating process is managed by macrophages. The increase in pro-inflammatory cytokine genes provides suppression of tumor cells in the environment. It was aimed to investigate the effect of Odoroside A on macrophage transformation. Materials and methods: The non-lethal dose of Odoroside A in macrophage cells was determined by CCK-8 analysis. Then, gene expression analysis of cytokines genes and surface markers in RAW264.7 cells was performed by using qRT-PCR. The protein levels of the cytokines that were significant in the gene expression analysis were examined using the ELISA method. Results: Odoroside A significantly increased the expression of IL1 and IL6 genes in RAW264.7 cells, while it significantly decreased the expression of IL10 gene. There was no significant change in IL4 and TGFβ gene expressions. IL1 and IL6 gene expressions, which had a statistically significant increase, were also found to increase significantly at the protein level. Conclusions: When all the data are evaluated together, it has been observed that Odoroside A increases the pro-inflammatory response in RAW264.7 cells, thus providing M1 type macrophage transformation. This suggests that Odoroside A may be effective in tumor elimination of the macrophage-mediated environment in the tumor niche.

Anahtar Kelimeler

• Odoroside A
• macrophage
• cytokines

RAW264.7 hücrelerinde Odoroside A'nın immün transformasyondaki rolü

Öz

Amaç: Makrofajlar bağışıklık sisteminin en önemli unsurlarından biridir ve inflamasyonda ikili rol oynamaktadır. Hem ortamda immün yanıtın oluşumunu hem de baskılanmasını düzenlemektedir. Kanser mikroçevresindeki bu sinyalleme, kanserin ilerlemesini etkiler. Bu kanseri teşvik eden veya kanseri ortadan kaldıran süreç makrofajlar tarafından yönetilmektedir. Proinflamatuar sitokin genlerinin ifadesindeki artış, ortamdaki tümör hücrelerinin baskılanmasını sağlar. Bu çalışmada Odoroside A'nın makrofaj dönüşümüne etkisinin araştırılması amaçlanmıştır. Gereç ve yöntem: Makrofaj hücrelerinde öldürücü olmayan Odoroside A dozu CCK-8 analizi ile belirlenmiştir. Daha sonra RAW264.7 hücrelerinde sitokin genlerinin ve yüzey belirteçlerinin qRT-PCR kullanılarak gen ekspresyon analizi yapılmıştır. Gen ekspresyonu analizinde anlamlı çıkan sitokinlerin protein düzeyleri ELISA yöntemi kullanılarak incelenmiştir. Bulgular: Odoroside A, RAW264.7 hücrelerinde IL1 ve IL6 genlerinin ekspresyonunu önemli ölçüde artırırken, IL10 geninin ekspresyonunu önemli ölçüde azaltmıştır. IL4 ve TGFβ gen ifadelerinde anlamlı bir değişiklik olmamıştır. İstatistiksel olarak anlamlı artış gösteren IL1 ve IL6 gen ekspresyonlarının protein düzeyinde de anlamlı düzeyde arttığı belirlenmiştir. Sonuç: Tüm veriler birlikte değerlendirildiğinde Odoroside A'nın RAW264.7 hücrelerinde proinflamatuar yanıtı arttırdığı, dolayısıyla M1 tip makrofaj dönüşümünü sağladığı gözlenmiştir. Bu, Odoroside A'nın, tümör nişindeki makrofaj aracılı oluşan proinflamatuar ortamın tümörün ortadan kaldırılmasında etkili olabileceğini düşündürmektedir.

Anahtar Kelimeler

• Odoroside A
• makrofaj
• sitokinler

Kaynakça

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