Papain Induces Apoptosis by Regulating Caspase-3 and Caspase-9 Genes in HepG2 Cells

Öz

Hepatocellular carcinoma (HCC) ranks second in cancer-related death and its incidence is increasing worldwide. Papain is a proteolytic enzyme and a potential anticancer agent. The aim of the study is to evaluate the cytotoxic effect of papain in human hepatoma HepG2 cell line through apoptosis. The cytotoxicity of papain was determined by the MTT method. Morphological changes of Papain-treated HepG2 cells were evaluated by acridine orange and ethidium bromide (AO/EB) dual staining. Apoptotic activity was determined by the expression of apoptosis regulator caspase-3 and caspase-9 genes by qPCR method. As a result of the administration of 500 µg/ml, 250 µg/ml, 100 µg/ml, 50 µg/ml, 25 µg/ml and 10 µg/ml papain to HepG2 cells for 48 hours, the IC50 value was found to be 53 µg/ml. Caspase 3 and 9 gene expressions, which are apoptotic markers, were significantly increased as a result of papain administration in HepG2 cells. In conclusion, papain may have anticancer effect for HCC treatment by inducing expression of apoptosis regulatory genes.

Anahtar Kelimeler

• Papain; HepG2; Hepatocellular carcinoma; Cytotoxicity; Apoptosis.

Papain HepG2 Hücrelerinde Kaspaz-3 ve Kaspaz-9 Genlerini Düzenleyerek Apoptozu İndükler

Öz

Hepatosellüler karsinom (HCC), kansere bağlı ölüm sıralamasında ikinci sırada yer alır ve dünya genelinde sıklığı artmaktadır. Papain proteolitik bir enzimdir ve potansiyel antikanser ajanıdır. Çalışmanın amacı, papainin insan hepatom HepG2 hücre hattındaki sitotoksik etkisinin apoptoz üzerinden değerlendirilmesidir. Papainin sitotoksisitesi MTT yöntemi ile belirlendi. Papain uygulanan HepG2 hücrelerinin morfolojik değişiklikleri, akridin portakalı ve etidyum bromür (AO/EB) ikili boyaması ile değerlendirildi. Apoptotik aktivite qPCR yöntemi ile apoptoz düzenleyici kaspaz-3 ve kaspaz-9 genlerinin anlatımları ile tespit edildi. HepG2 hücrelerine 48 saat boyunca 500 µg/ml, 250 µg/ml, 100 µg/ml, 50 µg/ml, 25 µg/ml ve 10 µg/ml papain uygulaması sonucunda IC50 değeri 53 µg/ml bulundu. Apoptotik belirteçler olan kaspaz 3 ve 9 gen ifadeleri, HepG2 hücrelerinde papain uygulaması sonucunda önemli ölçüde arttı. Sonuç olarak, papain HCC tedavisi için, apoptoz düzenleyici genlerin anlatımını indükleyerek antikanser etkiye sahip olabilir.

Anahtar Kelimeler

• Papain
• HepG2
• Hepatosellüler karsinom
• Sitotoksisite
• Apoptoz

Kaynakça

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