Meme kanserinde maslinik asit ve oleanolik asit kombinasyonunun wnt sinyal yolağı üzerindeki etkilerinin moleküler düzeyde araştırılması
Küçük Resim Yok
Tarih
2022
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Tüm dünyada kadınları etkileyen ve oldukça yaygınlaşan meme kanseri, büyük ölçüde heterojenite gösteren karmaşık bir hastalık olduğu için tedavi modellerinde kişiye özel bir yaklaşım gereklidir. Oleaceae familyasına ait bitkilerde doğal olarak oluşan bir pentasiklik triterpenoid bileşiği olan Maslinik asit ve Oleanolik asidin anti-kanser etkisi çok sayıda klinik öncesi deneyle doğrulanmıştır. Wnt/?-katenin sinyal yolunun meme kanserinde aktive olması kadınlarda kanser ölümlerinin önde gelen nedenidir. Bu yüzden anormal Wnt aktivasyonunun moleküler mekanizmalarını belirlemek, yolak hedefli tedavinin geliştirilmesi için kritik öneme sahiptir. Tez çalışmasında meme kanseri hücre hatlarında kullanılan doğal bileşiklerin sitotoksik etkisi ve WNT sinyal yolağı ile ilişkili gen ekspresyon değişimlerinin aydınlatılması hedeflenmiştir. Bu amaçla maslinik asit ve oleanolik asidin tek tek ve kombinasyon olarak uygulanmasının insan meme kanseri hücre hattı MCF-7 üzerindeki sitotoksisitesi WST-1 testi ile belirlenmiştir. Ayrıca MCF-7 hücrelerinde maslinik asit, oleanolik asit ve MA+OA kombinasyon uygulaması ile WNT sinyal yolağı ile ilişkilendirilmiş olan genlerde meydana gelen ekspresyon değişiklikleri qRT-PCR ile tespit edilmiştir. Çalışmada maslinik asidin MCF-7 hücreleri için IC50 değeri 72 saatte 70,3 ?M, oleanolik asidin IC50 değeri 72 saatte 291 ?M ve kombinasyonun ED50 konsantrasyonları, MA için 3.45 DRI ve 20.34 ?M, OA için 3.09 DRI ve 94.01µM, kombinasyon indeksleri (CI) 0.612 sinerjizim olarak saptanmıştır. Maslinik asit ve Oleanolik asidin apoptoz ve otofajiyi indüklediği ve aynı zamanda G0/G1 hücre siklüsü arrestine yol açtığı belirlenmiştir. WNT sinyal yolağı ile ilişkili genlerin ekspresyon düzeyleri incelendiğinde BTRC, CCND2, CTNNBIP1, EP300, FBXW4, FOXN1, FZD9, GSK3A, GSK3B, LEF1, MAKP8, NLK genlerinin ekspresyon seviyelerinde artış; AXIN2, PRICKLE1, WNT3, WNT5A, WNT5B, WNT6, WNT7B genlerinin ekspresyon seviyelerinde ise azalma tespit edilmiştir. Çalışmamızdan elde edilen sonuçlara bakıldığında WNT yolağındaki tümör supresör genlerin upregüle olması ve WNT sinyalini indükleyen onkogenlerin downregüle olduğunun belirlenmesi ile meme kanseri patogenezinin daha iyi anlaşılması için literatüre katkı sağlayacaktır. MA+OA kombinasyonu kanser hücrelerinde apoptoza neden olmuştur. Doğal bileşiklerin kombinasyon olarak kullanımı kanser hücrelerinin tedaviye direncini azaltabilir. Böylece kullanılan doğal ve bitkisel ürünlerin kombinasyonunun meme kanseri tedavisini destekleyebilecek umut verici yeni bir hedef molekül olabilme ihtimali ortaya konmuştur
Breast cancer, which affects women all over the world and has become very common, is a complex disease with a large heterogeneity. For this reason, a personalized approach is required in treatment models. The anti-cancer effect of Maslinic acid and Oleanolic acid, a naturally occurring pentacyclic triterpenoid compound in plants belonging to the Oleaceae family, has been confirmed by numerous preclinical experiments. Activation of the Wnt/?-catenin signaling pathway in breast cancer is the leading cause of cancer death in women. Therefore, identifying the molecular mechanisms of abnormal Wnt activation is critical for the development of pathway-targeted therapy. In the thesis study, it was aimed to elucidate the cytotoxic effect of natural compounds used in breast cancer cell lines and the gene expression changes associated with the WNT signaling pathway. For this purpose, the cytotoxicity of the treatment of maslinic acid and oleanolic acid individually or in combination on the human breast cancer cell line MCF-7 was determined by the WST-1 test. In addition, expression changes in genes associated with WNT signaling pathway were detected by qRT-PCR in MCF-7 cells with maslinic acid, oleanolic acid and MA+OA combination treatment. In the study, the IC50 value of maslinic acid for MCF-7 cells was found to be 70.3 ?M in 72 hours, and the IC50 value of oleanolic acid was 291 ?M in 72 hours. The ED50 concentrations of the combination were determined as 3.45 DRI and 20.34 µM for MA, 3.09 DRI and 94.01 µM for OA, and combination indexes (CI) 0.612 synergism. Maslinic acid and Oleanolic acid were found to induce apoptosis and autophagy and also cause G0/G1 cell cycle arrest. When the expression levels of genes associated with the WNT signaling pathway are examined, the expression levels of BTRC, CCND2, CTNNBIP1, EP300, FBXW4, FOXN1, FZD9, GSK3A, GSK3B, LEF1, MAKP8, NLK genes increased; the expression levels of AXIN2, PRICKLE1, WNT3, WNT5A, WNT5B, WNT6, WNT7B genes were decreased. Considering the results obtained from our study, the determination of the upregulation of tumor suppressor genes in the WNT pathway and the downregulation of oncogenes that induce WNT signaling will contribute to the literature for a better understanding of the pathogenesis of breast cancer. The combination of MA+OA caused apoptosis in cancer cells. The use of natural compounds in combination can reduce the resistance of cancer cells to treatment. Thus, the possibility that the combination of natural and herbal products used could be a promising new target molecule that can support breast cancer treatment has been exhibit.
Breast cancer, which affects women all over the world and has become very common, is a complex disease with a large heterogeneity. For this reason, a personalized approach is required in treatment models. The anti-cancer effect of Maslinic acid and Oleanolic acid, a naturally occurring pentacyclic triterpenoid compound in plants belonging to the Oleaceae family, has been confirmed by numerous preclinical experiments. Activation of the Wnt/?-catenin signaling pathway in breast cancer is the leading cause of cancer death in women. Therefore, identifying the molecular mechanisms of abnormal Wnt activation is critical for the development of pathway-targeted therapy. In the thesis study, it was aimed to elucidate the cytotoxic effect of natural compounds used in breast cancer cell lines and the gene expression changes associated with the WNT signaling pathway. For this purpose, the cytotoxicity of the treatment of maslinic acid and oleanolic acid individually or in combination on the human breast cancer cell line MCF-7 was determined by the WST-1 test. In addition, expression changes in genes associated with WNT signaling pathway were detected by qRT-PCR in MCF-7 cells with maslinic acid, oleanolic acid and MA+OA combination treatment. In the study, the IC50 value of maslinic acid for MCF-7 cells was found to be 70.3 ?M in 72 hours, and the IC50 value of oleanolic acid was 291 ?M in 72 hours. The ED50 concentrations of the combination were determined as 3.45 DRI and 20.34 µM for MA, 3.09 DRI and 94.01 µM for OA, and combination indexes (CI) 0.612 synergism. Maslinic acid and Oleanolic acid were found to induce apoptosis and autophagy and also cause G0/G1 cell cycle arrest. When the expression levels of genes associated with the WNT signaling pathway are examined, the expression levels of BTRC, CCND2, CTNNBIP1, EP300, FBXW4, FOXN1, FZD9, GSK3A, GSK3B, LEF1, MAKP8, NLK genes increased; the expression levels of AXIN2, PRICKLE1, WNT3, WNT5A, WNT5B, WNT6, WNT7B genes were decreased. Considering the results obtained from our study, the determination of the upregulation of tumor suppressor genes in the WNT pathway and the downregulation of oncogenes that induce WNT signaling will contribute to the literature for a better understanding of the pathogenesis of breast cancer. The combination of MA+OA caused apoptosis in cancer cells. The use of natural compounds in combination can reduce the resistance of cancer cells to treatment. Thus, the possibility that the combination of natural and herbal products used could be a promising new target molecule that can support breast cancer treatment has been exhibit.
Açıklama
04.04.2023 tarihine kadar kullanımı yazar tarafından kısıtlanmıştır
Anahtar Kelimeler
Tıbbi Biyoloji, Medical Biology
