Gossypinin insan hepatom (Hep-3B) hücreleri üzerindeki anti-proliferatif etkisi

Year 2020, Volume: 45 Issue: 3, 1165 - 1172, 30.09.2020

İrfan Çınar , Muhammed Yayla Damla Binnetoğlu

https://doi.org/10.17826/cumj.732912

Cited By: 3

Abstract

Amaç: Çalışmamızda Hibiscus vitifolius’dan izole edilen ve antioksidan, antienflamatuar, analjezik ve anti kanser özelliklere sahip olan gossypinin Hep-3B hücreleri üzerindeki antiproliferatif etkilerini göstermeyi amaçladık.
Gereç ve Yöntem: Çalışmamızda Hep-3B hücre hatları American Type Culture Collection (ATCC, USA) temin edilmiştir. Hücreler farklı konsantrasyonlarda (5-100 µg/ml) gossypin ve pozitif kontrol olarak da sisplatine (50µM) maruz bırakılmışlardır. Sonrasında 24, 48 ve 72 saatlerde hücrelere (MTT) yöntemi ile canlılık analizi yapılmıştır. Hücrelerdeki apoptozun göstergesi için Hoechst floresan boyama yapıldı. Aynı zamanda RT-PCR ile Nuklear kappa B (NFκB), kaspaz 3 ve 9 mRNA ekspresyon düzeyleri incelendi.
Bulgular: Çalışmamızda gossypin uygulaması doza ve zamana bağlı olarak hücre proliferasyonunu önlemiştir. İlk 24 saatte sadece 100µg/ml dozunda etki gösterirken 48 ve 72. saatlerde ise doza bağlı olarak etkisini göstermiştir. Hoechst floresan boyama ile gossypin 50 ve 100 µg/ml dozunda hücreleri daha belirgin apoptoza götürdüğü görülürken sisplatin grubu ile neredeyse aynı etkiyi ortaya koymuştur. NFκB mRNA ekspresyonunu doza bağlı inhibe eden gossypin, aynı zamanda apoptotik protein olan kaspas 3 ve 9’un mRNA ekspresyonlarını doza bağlı olarak indüklemiştir.
Sonuç: Gossypin doza bağlı olarak Hep-3B hücreleri üzerinde sisplatin ile yakın bir etki ortaya koymuştur. Bu etkilerini ise apoptozu aktive ederek ve NFκB inhibisyonu yaparak ortaya koymuştur. Buradan yola çıkarak karaciğer kanserinin tedavisinde gossypinin gelecekte potansiyel bir antikanser ajan olabilir. 

Keywords

Gossipin, Kanser, Hepatom

Supporting Institution

Yok

Project Number

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Thanks

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Antiproliferative effect of gossypin on human hepatoma (Hep-3B) cells

Abstract

Purpose: The aim of this study was to demonstrate the antiproliferative effects of Gossypin which has antioxidant, anti-inflammatory, analgesic and anti-cancer properties on Hep-3B cells isolated from Hibiscus vitifolius.
Materials and Methods: Hep-3B Cell Lines American Type Culture Collection (ATCC, USA) was provided in our study. The cells were exposed to different concentrations (5-100 µg/ml) of gossypin and cisplatine (50µM) as positive control. Afterwards, viability analysis was performed with MTT method on the cells at 24, 48 and 72 hours. Hoechst fluorescent staining was performed for indication of apoptosis in cells. At the same time, RT-PCR and NFκB, caspase 3 and 9 mRNA expression levels were examined.
Results: Gossypin administration prevented cell proliferation due to dose and time. In the first 24 hours, only 100µg/ml dose was effective, while in 48 and 72 hours it was effective depending on the dose. With Hoechst fluorescent staining, gossypin showed nearly the same effect as the cisplatin group, with doses of 50 and 100 µg/ml seen to lead the cells to more apoptosis. Gossypin, which inhibits dose dependent NFκB mRNA expression, has also up regulated dose dependent the apoptotic proteins mRNA expression of caspase 3 and 9.
Conclusion: Gossypin showed a close effect with cisplatin on Hep-3B cells depending on the dose. These effects were demonstrated by activating apoptosis and inhibiting NFκB. Based on this, gossypin may be a potential anticancer agent in the future for the treatment of liver cancer.

Keywords

Gossipin, hepatom, cell line

Project Number

Yok

References

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