MEME KANSERİ VE HEPATOSELLÜLER KANSER HÜCRE DİZİLERİNDE AMPK MODÜLASYONUNUN KANSER HÜCRE PROLİFERASYONU ÜZERİNE ETKİSİNİN GERÇEK-ZAMANLI HÜCRE ANALİZ SİSTEMİ xCELLigence ARACILIĞIYLA İNCELENMESİ
AMP (adenozin monofosfat) aktive edici protein kinaz (AMPK), hücrenin çeşitli metabolik stres durumlarındaaktive olarak hücredeki enerji homeostazını sağlayan önemli bir serin / treonin protein kinazdır. Stres ve hipoksikortam koşulları gibi organizmadaki enerji miktarının düşük olduğu veya enerji tüketiminin yüksek olduğu koşullardahücresel AMP miktarı artmakta ve AMPK aktivasyonu ile birlikte yağ asidi oksidasyonu ve glikolizis gibi katabolikreaksiyonlar artmaktadır. AMPK aktivasyonu; birçok kanser türünde deregüle olmaktadır. Yapılan çeşitli çalışmalar,AMPK agonisti ajanlar ile kanser hücrelerinin hedeflenebileceğini ve bu sayede ortalama sağkalımın arttırılabileceğiniöne sürmektedir. Bu görüşün aksine bazı çalışmalar ise, tümör gelişimi sürecinde AMPK aktivasyonunun arttığınıgöstermektedir. Kanser hücrelerinin olumsuz koşullar altında AMPK aktivasyonu gösterdiği ve AMPK'ın protümöralözellikte olduğu iddia edilmektedir. Kanser hücreleri ile yapılan çalışmalarda ortaya çıkan bu çelişkili durumun,hücrenin moleküler profiline ve tümör mikroçevresindeki koşullara bağlı olduğu öne sürülmektedir. Ayrıca, AMPKaktivitesi enflamasyonu ve anti-tümör immün yanıtları düzenleyerek tümör gelişimi ve ilerlemesini azaltabilir.Bu çalışmada, farklı moleküler profillere sahip meme kanseri (SK-BR-3) ve hepatosellüler kanser (Huh-7) hücredizileri, AMPK inhibitör ve aktivatör ajanları ile ayrı ayrı olarak inkübe edilmiştir. İlaç inkübasyonları sonucuhücre proliferasyon paternleri, gerçek-zamanlı hücre analiz sistemi (xCELLigence) kullanılarak analiz edilmiş vegözlemlenen değişimler kendi aralarında ve ilaç ile inkübe edilmemiş kontrol gruplarıyla karşılaştırılmıştır. Busayede, iki farklı kanser türünde AMPK hedefine yönelik doğru stratejinin karşılaştırılmalı olarak belirlenmesihedeflenmiştir. Genel olarak AMPK aktivasyonu, hem meme kanserinde hem de hepatosellüler kanserde hücreproliferasyonunu ve agresifliğini arttırmıştır. Öbür taraftan, AMPK inhibisyonu ise genel anti-tümöral etkilergöstermiştir. Ancak, hepatosellüler kanser hücrelerinde düşük-doz AMPK inhibisyonuyla proliferasyon artmıştır.Kanser tedavisinde AMPK modülasyonunun, kanser metabolizması ve anti-tümör immün yanıtlar üzerinde kritikbir regülasyon potansiyeli olduğu unutulmamalıdır. Bu çalışmanın sonuçları, kanser metabolizmasını ve anti-tümörimmün yanıtları hedef alan anti-kanser tedavi stratejilerinin farklı kanserlerde dikkatle geliştirilmesi ve uygulanmasıgerekliliğine dikkat çekmektedir
Evaluation of the Effects of AMPK Modulation on Cancer Cell Proliferation via Real-Time Cell Analysis System xCELLigence in Breast and Hepatocellular Cancers
AMP (adenosine monophosphate) activating protein kinase (AMPK), is a crucial serine / threonine protein kinase that is activated in several cellular metabolic stress conditions in order to maintain cellular energy metabolism. In states of low energy or high energy expenditure in the organism such as conditions of stress and hypoxia, cellular AMP level increases and AMPK becomes activated. This, in turn, results in an increase in catabolic reactions like fatty acid oxidation and glycolysis. AMPK activation becomes dysregulated in several types of cancer. Several studies suggest that AMPK agonist agents can be utilized to target cancer cells, which may increase mean survival. On the contrary, several other studies report that AMPK activation is increased during tumor development. It is proposed that cancer cells show AMPK activation under unfavorable conditions and AMPK has pro-tumoral features. This contradiction that has arisen in studies with varying results obtained from cancer cells was proposed to be due to the molecular profile of the cells and the conditions of the tumor microenvironment. In addition, AMPK activity may decrease tumor development and progression by regulating inflammation and anti-tumor immune responses. In the current study, breast (SK-BR-3) and hepatocellular (Huh-7) cancer cell lines with different molecular profiles were incubated with either AMPK inhibitor or activator agents. Cellular proliferation patterns after drug incubations were analyzed with a real-time cell analysis system (xCELLigence) and the results were compared with other treatment group as well as the control group was not incubated with any of the drugs. By this way, we aimed to determine the right strategy to target AMPK in two different cancer types in a comparative manner. In general, activation of AMPK increased cancer cell proliferation and aggressiveness in both breast and hepatocellular cancers. On the other hand, AMPK demonstrated anti-tumoral effects in general. However, lowdose AMPK inhibition increased the proliferation of hepatocellular cancer cells. It should be kept in mind that AMPK modulation as a cancer treatment approach has a critical regulatory potential on cancer metabolism and anti-tumor immune responses. The results of the current study call attention to the importance of meticulous development and application of anticancer treatment modalities that target cancer metabolism in different cancer types.
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