Derya BABACAN, Sümeyye AYDOĞAN TÜRKOĞLU, Feray KÖÇKAR

Kolon Kanserinde IL-6 Sitokinin Karbonik anhidraz III Gen İfadesi Üzerindeki Etkisinin İncelenmesi

Yüksek IL-6 seviyeleri, birçok kanser türünde agresif tümör büyümesi ve tedaviye yanıt ile ilişkilidir. Dolaşımdaki IL-6 düzeyi yüksek olan hastalar genellikle kötü prognoz ve daha kısa sağkalım ile ilişkilendirilirken, daha düşük bir IL-6 düzeyi tedaviye daha iyi yanıt ile ilişkilidir. Kolorektal kanser (KRK)’in IL-6 ile regüle olduğu ve KRK hastalarında serum IL-6 seviyesinin arttığı, bu artışında tümör boyutuyla ilişkili olduğu gösterilmiştir. Organizmalarda uygun asit-baz dengesini sağlamada önemli rol oynayan hidrataz aktivitesine sahip olan CAIII bu görevinin dışında reaktif oksijen radikallerinin yok edilmesi ile oksidatif stresin neden olduğu hasardan hücreleri korumada, ayrıca glikolitik ara ürünlerinden oksaloasetatın ve sitratın hızlı dönüşümünü kolaylaştırarak bunların yağ asit metabolizmasına katılmasını da sağlamaktadır. Son yapılan çalışmalarda ise CAIII’ün ve kanser ilişkisi üzerinde durulmaktadır. Karaciğer karsinomda (HCC), CAIII ekspresyonunun önemli derecede azaldığı ve hücrelerin invasyonunu arttırdığı gösterilmiştir. Kolon kanserinde ise CAIII ifadesinin MEK-1 ve PI3K yolakları yoluyla baskılandığı da tespit edilmiştir. Prostat kanserinde de hipoksik yolun regülatör proteini olan HIF1-α'nın doğrudan CAIII promotoruna bağlandığı ve kanser sürecinde aktif rol oynadığı gösterilmiştir. Çalışmamızda IL-6 sitokinin CAIII geni üzerindeki regülasyonu 2 farklı basamakta değerlendirilmiştir. 500 U IL-6 sitokini uygulanan HT-29 hücrelerinde CAIII mRNA seviyesi Real Time PCR ile değerlendirilmiş ve yine 500 U IL-6 uygulanan HT-29 hücrelerinden elde edilen protein ekstraklarının kullanıldığı western blot tekniği ile CAIII protein seviyesi üzerindeki etkisi tespit edilmiştir. Sonuç olarak bu çalışmada IL-6 etkili CAIII regülasyonunda CAIII’ün hem mRNA hem de protein düzeyindeki ifadesinin, IL-6 muamelesi görmemiş kontrol grubu hücrelerine kıyasla azaldığı tespit edilmiştir. Buna göre; IL-6 sitokininin kanser ilgili bir protein olan CA3 üzerindeki düzenleyici etkisi olduğu tespit edilmiştir.

Anahtar Kelimeler:

Karbonik Anhidraz III, kolon kanseri, IL-6

Investigation of the Effect of IL-6 Cytokine on Carbonic Anhydrase III Gene Expression in Colon Cancer

High IL-6 levels are associated with aggressive tumor growth and response to therapy in many types of cancer. Patients with high circulating IL-6 levels are generally associated with poorer prognosis and shorter survival, while a lower IL-6 level is associated with better response to treatment. Studies conducted in recent years emphasize that colorectal cancer (CRC) is regulated by IL-6, serum IL-6 level is increased in CRC patients and this increase is correlated with tumor size. Having hydratase activity, which plays an important role in maintaining the proper acid-base balance in organisms, CA3 also protects cells from damage caused by oxidative stress by destroying reactive oxygen radicals, and also facilitates the rapid conversion of glycolytic intermediates oxaloacetate and citrate, enabling them to participate in fatty acid metabolism. Recent studies have focused on the relationship between CAIII and cancer. It has been determined that CAIII increases the invasion of cells and the expression of CAIII is significantly decreased in Liver carcinoma (HCC). It has also been determined that CAIII expression is suppressed through MEK-1 and PI3K pathways in colon cancer cell line HT-29 cells. Finally, it has been shown that HIF1-α, the regulatory protein of the hypoxic pathway in prostate cancer, directly binds to the CAIII promoter. In our study, the regulation of IL-6 cytokine on the CAIII gene was evaluated in 2 different steps. The effect of CAIII mRNA levels and protein levels were determined by Real Time PCR and Western blot analysis, respectively. The sample in which total RNAs and protein obtained from HT-29 cells treated with 500 U of IL-6 cytokine were used for both analysis. It was determined that the expression of CAIII at both mRNA and protein levels were decreased by the effective regulation of IL-6.In conclusion, this study elucidates the regulatory effect of IL-6 cytokine on cancer-related protein CA3 gene regulation.

Keywords:

Carbonic Anhydrase III, colon cancer, IL-6,

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