Yaprak DÖNMEZ ÇAKIL, İlayda ALTUN, Elif TEKİN İŞLEREL, Zeynep OZUNAL

Sertralin ve sorafenibin karaciğer kanseri hücrelerinde P-glikoprotein gen ekspresyonuna ve rodamin 123 birikimine etkileri

Amaç: Hepatoselüler karsinom (HCC), tüm dünyada görülme sıklığı giderek artan, en yaygın ölümcül kanser türleri arasında yer almaktadır. Vakaların büyük çoğunluğunda ileri evrede tanı ve ilaç direnci hayatta kalma oranını sınırlayan temel sorunlardır. ATP bağımlı bir efluks (dışa atım) pompası olan P-glikoprotein (P-gp), kanserde çok sayıda ilaç direnci modeli ile ilişkilidir. Rodamin 123 (rh123) floresan bir boyadır ve bir referans P-gp substratı olarak birçok araştırmada P-gp aktivitesini incelemek amacıyla kullanılmaktadır. Sorafenib, HCC tedavisi için onaylanmış ilk sistemik tedavidir. Etkinliğini artırmak ve ilaç direncini azaltmak için farklı ilaçlarla beraber kullanımları araştırılmaktadır. Bu kapsamda, daha önce sorafenib ve antidepresan sertralinin HepG2 hücre proliferasyonu ve ölümü üzerinde sinerjistik etkileri gösterilmiştir. Yöntem: Bu çalışmada, HepG2 hücrelerinde, sorafenib ve sertralinin tek başına veya birlikte uygulanması sonrası, qPCR ve akış sitometrisi ile P-gp gen ekspresyonu ve rh123 birikimi/efluks araştırılmıştır. Bulgular: Sorafenib uygulaması hem P-gp gen ekspresyonu hem de hücrelerde rh123 birikimini anlamlı olarak azaltırken, sertralin tek başına kullanıldığında P-gp mRNA seviyelerini arttırmış, rh123 birikiminde ise anlamlı bir değişikliğe yol açmamıştır. İki ilacın birlikte uygulanması ise, kontrol grubuna kıyasla P-gp gen ekspresyonunda ve rh123 birikiminde herhangi bir değişikliğe neden olmamıştır. Sonuç: Sonuçlar, sorafenib ve sertralinin antiproliferatif sinerjistik etkilerinde P-gp ile ilişkili etkenlerden ziyade farklı mekanizmaların rol alabileceğini göstermiştir.

Anahtar Kelimeler:

Sertraline, sorafenib, sinerjist etki, P-glikoprotein, HepG2, rodamin 123

The effects of sertraline and sorafenib on P-glycoprotein expression and rhodamine 123 accumulation in liver cancer cells

Aim: Hepatocellular carcinoma (HCC) is among the most common lethal cancer type with an increasing incidence all over the world. In the majority of cases, advanced stage diagnosis and drug resistance are the main issues that limit the survival rate. P-glycoprotein (P-gp), an ATP-dependent efflux pump, is associated with numerous drug resistance patterns in cancer. Rhodamine 123 (rh123) is a fluorescent dye used as a reference substrate for measurement of P-gp activity in various studies. Sorafenib is the first approved systemic therapy for HCC treatment. Novel combined approaches of sorafenib with cytotoxic drugs are extensively studied to increase its effectiveness and overcome drug resistance. Recently, the synergistic effects of sorafenib and antidepressant sertraline on HepG2 cell proliferation and death were shown. Method: In the present study, we investigated the possible effects of sorafenib and sertraline as single agents or in combination on P-gp expression and rh123 accumulation/efflux by qPCR and flow cytometry, respectively. Results: Sorafenib administration significantly decreased both P-gp gene expression and rh123 accumulation in cells, while sertraline increased P-gp mRNA levels, it did not cause a significant change in rh123 accumulation. Co-administration of the two drugs did not cause any alterations in P-gp gene expression and rh123 accumulation compared to the control group. Conclusion: The results suggested that the presence of other mechanisms rather than P-gp associated effects are responsible for the synergistic activity of sorafenib and sertraline.

Keywords:

Sertraline, sorafenib, synergy, p-glycoprotein, HepG2, rhodamine123,

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