Publication: Glycyrrhiza glabra l. ekstresinin meme kanseri hücrelerinde dosetaksel ile kombine tedavide antiproliferatif etkisi ve bu etkide protein disülfit izomerazın rolü
Abstract
Glycyrrhiza glabra l. ekstresinin meme kanseri hücrelerinde dosetaksel ile kombine tedavide antiproliferatif etkisi ve bu etkide protein disülfit izomerazın rolü Amaç: Bu çalışmada, Glycyrrhiza glabra L.’nin dosetaksel ile kombine kullanımında dosetakselin sitotoksik rolü ile ilaç direncine etkisini ve G.glabra‘nın meme kanseri hücreleri üzerindeki sitotoksik etkisinde protein disülfit izomeraz (PDI) inhibisyonu üzerindeki etkisini ortaya koymak amaçlanmıştır. Gereç ve Yöntem: G. glabra L. bitkisinin iki farklı varyetesine ait olmak üzere toplam 3 kök droğun etanollü (%80) ekstrelerinin; antioksidan aktiviteleri, total fenolik (TFM), flavonoid (TFLM) triterpen (TTM) madde miktarları belirlendi ve içerik analizleri yapıldı. Hazırlanan ekstrelerin ve dosetakselin MTT ile MCF-7 hücrelerinde canlılık üzerine etkilerine bakıldı. Sitotoksik etkisi en yüksek bulunan ekstrenin (GF-1) dosetaksel ile değişen konsantrasyonlarda kombine kullanımında sinerjik etkileri kombinasyon indeksi ile belirlendi. Kombine tedavinin hücre ölüm yollarına etkisi Anneksin V-PI boyama, kaspaz 3 aktivitesi ile belirlendi. Etkin ekstrenin PDI inhibisyonuna etkisine bakıldı. İçerik analizi ile belirlenen majör bileşenlerin PDI ile moleküler kenetleme çalışmaları yapıldı. Bulgular: Ekstrelerin ve dosetakselin anlamlı bir şekilde (p <0,05) hücre canlılığını azalttığı bulundu. G.glabra var. glandulifera (GF-1) ekstresinin NIH-3T3 sağlıklı hücrelerde sitotoksik etkisi gözlenmedi (p<0,01). 24 saatlik uygulamada, dosetakselin 62,5 µg/ ml konsantrasyonu ile; 48 saatlik uygulamada dosetakselin 240 µg/ ml ve 60 µg/ ml konsantasyonları ile GF-1 in tüm dozları sinerjik etki gösterdi. Kombine tedavide MCF-7 hücrelerinde canlılık ve erken apoptoz düzeylerinde ve kaspaz 3 aktivitesinde diğer gruplara göre anlamlı farklılık bulundu (p<0.05). GF-1’in konsantrasyona bağlı olarak PDI aktivitesini baskıladığı görüldü. Moleküler kenetleme çalışmalarında D-pinitol, glisirizik asit ve floretik asidin PDI ile etkileşiminde RMSD değerlerinin <2 AO olduğu bulundu. Sonuç: G.glabra, MCF-7 meme kanseri hücrelerinde kombine kullanımda dosetakselin düşük dozlarda kullanımında etkinliğini arttırarak kemoterapinin toksik yan etkilerini azaltabilir ve kemorezistans durumunda tedavinin etkinliğini arttırabilir. Bu etkisinde PDI’nın inhibisyonunun rolü olabilir.
Antiproliferative effect of glycyrrhiza glabra l extract in combination treatment with docetaxel in breast cancer cells and the role of protein disulfide isomerase in this effect Objective: We aimed to investigate the effect of Glycyrrhiza glabra to cytotoxic role and kemoresistance of docetaxel in combination with docetaxel and to reveal the role of protein disulfide isomerase (PDI) inhibition in the cytotoxic effect of G. glabra on breast cancer cells. Materials and Methods: Antioxidant activities, total phenolic (TFM), flavonoid (TFLM) and triterpene (TTM) content of ethanolic (80%) extracts of the three root from two different varieties of G.glabra L. were determined and content analyses were performed. The effects of the extracts and docetaxel on viability in MCF-7 cells with MTT were examined. The synergistic effects of the extract with the highest cytotoxic effect (GF-1) in combination with docetaxel at varying concentrations were determined by combination index. The effect of the combined treatment on cell death pathways was determined by Annexin V-PI and caspase 3 activity. The effect of the active extract on PDI inhibition was examined. Molecular docking studies were performed with PDI for the major components determined by content analysis. Results: It was found that extracts and docetaxel significantly (p< 0.05) decreased cell viability. No cytotoxic effect of G.glabra var. glandulifera (GF-1) extract was observed on NIH-3T3 healthy cells (p < 0.01). In 24-hours, docetaxel with 62.5 µg/ ml concentration; in 48-hours, docetaxel with 240 µg/ ml and 60 µg/ ml concentrations and all doses of GF-1 showed synergistic effect. In combined treatment, significant differences were found in viability and early apoptosis levels and caspase 3 activity in MCF-7 cells compared to the other groups (p<0.05). GF-1 was observed to suppress PDI activity depending on the concentration. In molecular docking studies, RMSD values were found to be <2 AO in the interaction of D-pinitol, glycyrrhizic acid and phloretic acid with PDI. Conclusion: G.glabra L. may reduce the toxic side effects of chemotherapy by increasing the efficacy of low-dose docetaxel in combined use in MCF-7 breast cancer cells and may increase the efficacy of treatment in cases of chemoresistance. Inhibition of PDI may play a role in this effect.
Antiproliferative effect of glycyrrhiza glabra l extract in combination treatment with docetaxel in breast cancer cells and the role of protein disulfide isomerase in this effect Objective: We aimed to investigate the effect of Glycyrrhiza glabra to cytotoxic role and kemoresistance of docetaxel in combination with docetaxel and to reveal the role of protein disulfide isomerase (PDI) inhibition in the cytotoxic effect of G. glabra on breast cancer cells. Materials and Methods: Antioxidant activities, total phenolic (TFM), flavonoid (TFLM) and triterpene (TTM) content of ethanolic (80%) extracts of the three root from two different varieties of G.glabra L. were determined and content analyses were performed. The effects of the extracts and docetaxel on viability in MCF-7 cells with MTT were examined. The synergistic effects of the extract with the highest cytotoxic effect (GF-1) in combination with docetaxel at varying concentrations were determined by combination index. The effect of the combined treatment on cell death pathways was determined by Annexin V-PI and caspase 3 activity. The effect of the active extract on PDI inhibition was examined. Molecular docking studies were performed with PDI for the major components determined by content analysis. Results: It was found that extracts and docetaxel significantly (p< 0.05) decreased cell viability. No cytotoxic effect of G.glabra var. glandulifera (GF-1) extract was observed on NIH-3T3 healthy cells (p < 0.01). In 24-hours, docetaxel with 62.5 µg/ ml concentration; in 48-hours, docetaxel with 240 µg/ ml and 60 µg/ ml concentrations and all doses of GF-1 showed synergistic effect. In combined treatment, significant differences were found in viability and early apoptosis levels and caspase 3 activity in MCF-7 cells compared to the other groups (p<0.05). GF-1 was observed to suppress PDI activity depending on the concentration. In molecular docking studies, RMSD values were found to be <2 AO in the interaction of D-pinitol, glycyrrhizic acid and phloretic acid with PDI. Conclusion: G.glabra L. may reduce the toxic side effects of chemotherapy by increasing the efficacy of low-dose docetaxel in combined use in MCF-7 breast cancer cells and may increase the efficacy of treatment in cases of chemoresistance. Inhibition of PDI may play a role in this effect.