Fatma FIRAT, Canan TÜRKOĞLU, H. Seda VATANSEVER, Feyzan ÖZDAL KURT

Kolon Kanseri Kök Hücrelerinde Akteosid’in Inflamasyon yada Apoptoza Etkisi Var mıdır?

Amaç: Kolon kanseri dünya çapında sağlık problemlerinin başında gelmektedir. Kanser kök hücreleri (CSC’ler), tümör heterojenitesi ve dormansisinde yer alan tümör başlatan hücreler olarak adlandırılır. CSC’ler ilaç direncine, metastaza ve primer ve metastatik kanserlerin nüksetmesine neden olabilir. Kolon kanser kök hücreleri etkili tedavi için alternatif bir yol olabilir. Çalışmamızda non-metastatik (HCT-116) ve metastatik (Colo-741) kolon kanseri hücreleri ve CSC’lerinde, akteosidin kök hücre özelliği, apoptotik ve inflamatuar sürece etkilerini değerlendirmeyi amaçladık. Gereç ve Yöntem: CSC’ ler, anti-CD133 belirteci kullanılarak MINIMACS sistemi ile her iki tip kolon kanseri hücre hattından elde edildi. Oct-4 ve CD 133 ekspresyonlarına bakılarak, metastatik COLO-741 ve non-metastatik HCT-116 CD133+ kanser kök hücreleri ve CD133- kanser hücreleri, 48 saat boyunca akteosid olmadan veya akteosid ile kültüre edildi. Apoptoz için Kaspaz-3, Bcl- 2, Bax, ve Fas-L, inflamasyon için IL-1β, TNF-α, IL-6, IL8 ve IL-10 proteinlerinin immunohistokimyasal ifadesi H-Score yapılarak istatistiksel olarak analiz edildi. Hücrelerin kök hücre özellikleri ve hücre morfolojisindeki değişiklikler uygulama öncesi ve sonrası karşılaştırmalı olarak belirlendi. Bulgular ve Sonuç: Oct-4 ve CD 133 ifadesinin hem metastatik hem de metastatik olmayan kolon kanser hücrelerinde akteosid uygulamasından sonra anlamlı derecede azaldığı görüldü. Ayrıca COLO-741 CD133+ ve HTC-116 CD133+ hücrelerin her ikisinde de Bax/Bcl oranının değiştiği görüldü. Akteosidin hücrelerde inflamasyona immunohistokimyasal olarak etki etmediği tespit edildi. Akteosidin kolon kanseri kök hücrelerinin kök hücre özelliğini ve apoptotik sürecini etkileyebileceği görülmüştür.

Does Acteoside Have Effects on Colon Cancer Stem Cells Via Inflamation or Apoptosis?

Aim: Colon cancer is one of the leading health problems worldwide. Cancer stem cells (CSCs) are referred to as tumor-initiating cells involved in tumor heterogeneity and dormancy. CSCs can cause drug resistance, metastasis, and recurrence of primary and metastatic cancers. Colon cancer stem cells may be an alternative route for effective treatment. In our study, we aimed to evaluate the effects of acteoside on stem cell properties, apoptotic and inflammatory processes in non-metastatic (HCT-116) and metastatic (Colo-741) colon cancer cells and CSCs. Materials and Methods: CSCs were obtained from both types of colon cancer cell lines with the MINIMACS system using the anti-CD133 reagent. After Oct-4 and CD 133 expressions specified, metastatic COLO-741 and non-metastatic HCT-116 CD133+ cancer stem cells and CD133- cancer cells were cultured with or without Acteoside for 48 hours. Immunohistochemical expression of Caspase-3, Bcl-2, Bax, and Fas-L for apoptosis, and IL-1β, TNF-α, IL-6, IL8 and IL-10 proteins for inflammation were statistically analyzed by performing H-Score. Changes in stem cell properties and cell morphology of the cells were determined comparatively before and after the application. Results and Conclusion: Expression of Oct-4 and CD 133 significantly decreased after actoside administration in both metastatic and non-metastatic colon cancer cells. In addition, it was observed that the Bax/Bcl ratio changed in both COLO-741 CD133+ and HTC-116 CD133+ cells. It was determined that acteoside did not affect inflammation in cells immunohistochemically. It has been observed that acteoside can affect the stem cell properties and apoptotic process of colon cancer stem cells.

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