Silibinin İn Vitro Ortamda CCL-228-SW 480 Kolon Kanseri Hücre Hattı Üzerine Apoptotik Etkisinin Araştırılması

Abstract

Amaç: Kolon kanseri, dünyada önemli bir morbidite ve mortalite nedenidir. Kolon kanseri, kansere bağlı ölümlerde erkeklerde akciğer ve prostat kanseri; kadınlarda meme ve akciğer kanserinden sonra üçüncü sırada yer almaktadır. Silibin, devedikeni (Silybum marianum) bitkisinden elde edilen bir ekstraktır. Yapılan çalışmalarda silibinin  deri, mide kanserlerinde kanser kemoprotektif ajan olduğu ve antikarsinojenik etki gösterdiği tespit edilmiş ayrıca meme, kolon, mesane kanseri hücre hatlarında hücre siklusunu durdurduğu ve apopitozisi tetiklediği akciğer tümörü büyümesini baskıladığı, gösterilmiştir. Çalışmamızda silibinin CCL-228-SW 480 kolon karsinoma hücre soyu üzerine etkisini monolayer kültür ortamında incelemeyi amaçladık.

Gereç Yöntem: Çalışmamızda silibinin CCL-228-SW 480 kolon karsinoma hücre soyu üzerine etkisini incelemek amacıyla 100 µM/ml doz silibin (ID50 inhibisyon dozu belirlenen)  hücrelere uygulandı. Silibin (100 µM/ml) uygulandıktan sonra  24, 48 ve 72 saat süreler sonunda BrdU işaretleme indeksi ve aktif kaspaz-3 tayinine immünohistokimyasal yöntemle bakıldı.

Bulgular: Sonuç olarak silibinin iki boyutlu kültür ortamında BrdU ile işaretlenen hücrelerin sayısını azalttığı, kaspaz-3 ile boyanan hücrelerin sayısını ise arttırdığı görüldü. Buradan da anlaşılacağı üzere silibin CCL-228-SW 480 kolon kanseri hücre soyunda hücre proliferasyonunu inhibe edici ve apoptozisi tetikleyici etki göstermiştir. Buradan yola çıkarak yapmış olduğumuz iki boyutlu kültür sonuçlarımızın daha sonra yapılacak silibin ve kanser çalışmalarına ışık tutacaği kanısındayız.

Keywords

• CCL-228-SW 480 hücre soyu,apopitozis,silibin

Investigation of Appoptotic Effect of Silibinin on CCL-228-SW 480 Column Cancer Cell Line: An In Vitro Study

Abstract

Aim: Colon cancer is a major cause of morbidity and mortality in the world.  It is third of cancer-related deaths after breast and lung cancer in women, lung cancer and prostate cancer in males. Silibin is an extract obtained from the plant of the thistle (Silybum marianum). Studies have shown that silibin is a cancer chemoprotective agent in skin, stomach cancers, and shows anticarcinogenic effect.At he same time it’s known tahat ıt suppresses cell cycle in breast, colon, bladder cancer cell lines and suppresses apoptosis-induced lung tumor growth. In our study, we aimed to investigate the effect of silibin on CCL-228-SW 480 colon carcinoma cell line in monolayer culture medium.       

Material and method: ID50 inhibition was determined on the dose for silibin and after ıt  was found 100 μM / ml was applied to the cells to examine the effect of silibin on CCL-228-SW 480 colon carcinoma cell line. Silibin (100 μM / ml) was administered and after 24, 48 and 72 hours the BrdU marking index and active caspase-3 assay were determined by immunohistochemistry.

Results: It was seen as a result silibin decreased the number of cells marked with BrdU in the two-dimensional culture medium and increased the number of cells stained with caspase-3. So, silibin CCL-228-SW480 inhibited cell proliferation and apoptosis in colon cancer cell line. We believe that the results of our two-dimensional culture will shed light on the silibin and cancer studies to be done later.

Keywords

• CCL-228-SW 480 cell line,apoptosis,silibin

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