Publication: KKeçi sütünden elde edilen protein hidrolizatlarının MCF-7 meme kanseri hücreleri üzerine etkilerinin incelenmesi
Abstract
Amaç: Bu çalışmanın amacı; protein parçalandığında aktif hale geçen ve insansağlığı üzerinde birçok yararlı etkileri olduğu bilinen biyoaktif peptitlerin MCF-7meme kanseri hücre dizileri üzerine olan etkilerini incelemektir.Gereç ve Yöntem: Bu çalışmada keçi sütünün yapısal özellikleri pH, karbohidrat,yağ, protein, nem, kuru madde ve kül açısından incelenmiştir. Keçi sütü kazein veserum fraksiyonlarına ayrıldıktan sonra pepsin, papain ve tripsin enzimleri ile ayrıayrı muamele ederek enzimatik olarak parçalanarak süt peptitleri elde edildi. Bupeptit molekülleri MCF-7 hücre dizileri ile 24 saat ve 48 saat farklıkonsantrasyonlarda inkübe edildi. İnkübasyon sonrası MCF-7 hücrelerinin canlılıkoranları 3-[4,5-dimetiltiazol-2-il]-2,5-difenil tetrazoliyum bromür canlılık testiyapılarak tayin edildi. Ölen MCF-7 hücrelerinden proteinler izole edildikten sonrakütle spektroskopisi sisteminde kütlelerine göre ayrılan proteinler ve peptitler tek tekkarakterize edildi.Bulgular: Keçi sütü serum protein fraksiyonunun pepsin ile muamelesinden sonra13 adet peptit, papain ile muamelesinden sonra 5 adet peptit ve tripsin ilemuamelesinden sonra 7-23 amino asit içeren, 39 adet peptit tespit edilmiştir. Kazeinfraksiyonunun pepsin ile muamelesinden sonra ise 10 adet peptit; papain ilemuamelesinden sonra 23 adet peptit ve tripsin ile muamelesinden sonra 21 adetpeptit tespit edilmiştir. MCF-7 hücrelerinin en yüksek apoptotik ölüm oranı kazeinfraksiyonunun pepsin ile muamelesinden sonra elde edilen peptidler ile olmuştur.Ölen hücrelerdeki proteinler karakterize edildiğinde hücre enerjisinin sağlanmasındayer alan proteinler ile ilişkili olan peptid dizileri tespit edilmiştir.Sonuç: Keçi sütü proteinlerinden elde edilen peptitlerin, yeni meme kanseri önleyiciilaç gelişimi için umut verici moleküller olduğu göz önünde bulundurulmalıdır.
Investigation of the Effects of Protein Hydrolysates Obtained from Goat Milkon MCF-7 Breast Cancer CellsName of students: Kıymet Özlem ŞAHNADepartment: Department of Biochemistry, Faculty of PharmacySupervisor: Prof. Dr. Tuğba Tunalı AKBAYObjective: The aim of this study was to investigate the effects of bioactive peptideson MCF-7 breast cancer cell lines, which are known to have many beneficial effectson human health when the protein breaks down.Materials and Methods: In the present study, the structural properties of goat milkwere examined in terms of pH, carbohydrate, fat, protein, moisture, dry matter andash. Goat milk was separated into casein and serum fractions and treated with pepsin,papain and trypsin enzymes. These peptide molecules were incubated with MCF-7cell lines at different concentrations for 24 hours and 48 hours. The viability rates ofMCF-7 cells after incubation were determined with 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide viability assay. After isolating proteins from MCF7 cells, proteins and peptides separated by mass in the mass spectroscopy systemwere individually characterized.Results: After treatment of goat milk serum protein fraction with pepsin, 13 peptide;after papain treatment 5 peptides and after treatment with trypsin 39 peptides weredetected. After treatment of the casein fraction with pepsin, 10 peptides; aftertreatment with papain, 23 peptides; after treatment with tripsin 23 peptides weredetected. The highest apoptotic mortality of MCF-7 cells was determined withpeptides which were obtained after the treatment of casein fraction with pepsin.When the proteins in the dead cells are characterized, the peptide sequencesassociated with the proteins involved in the delivery of the cell energy have beenidentified.Conclusion: It should be noted that peptides derived from goat milk proteins arepromising molecules for the development of new breast cancer-inhibiting drugs.
Investigation of the Effects of Protein Hydrolysates Obtained from Goat Milkon MCF-7 Breast Cancer CellsName of students: Kıymet Özlem ŞAHNADepartment: Department of Biochemistry, Faculty of PharmacySupervisor: Prof. Dr. Tuğba Tunalı AKBAYObjective: The aim of this study was to investigate the effects of bioactive peptideson MCF-7 breast cancer cell lines, which are known to have many beneficial effectson human health when the protein breaks down.Materials and Methods: In the present study, the structural properties of goat milkwere examined in terms of pH, carbohydrate, fat, protein, moisture, dry matter andash. Goat milk was separated into casein and serum fractions and treated with pepsin,papain and trypsin enzymes. These peptide molecules were incubated with MCF-7cell lines at different concentrations for 24 hours and 48 hours. The viability rates ofMCF-7 cells after incubation were determined with 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide viability assay. After isolating proteins from MCF7 cells, proteins and peptides separated by mass in the mass spectroscopy systemwere individually characterized.Results: After treatment of goat milk serum protein fraction with pepsin, 13 peptide;after papain treatment 5 peptides and after treatment with trypsin 39 peptides weredetected. After treatment of the casein fraction with pepsin, 10 peptides; aftertreatment with papain, 23 peptides; after treatment with tripsin 23 peptides weredetected. The highest apoptotic mortality of MCF-7 cells was determined withpeptides which were obtained after the treatment of casein fraction with pepsin.When the proteins in the dead cells are characterized, the peptide sequencesassociated with the proteins involved in the delivery of the cell energy have beenidentified.Conclusion: It should be noted that peptides derived from goat milk proteins arepromising molecules for the development of new breast cancer-inhibiting drugs.