The Synergistic Effect And Mechanism Of Ellagic Acid And GDC-0941 In Human Breast Cancer Treatment

Objective:The breast cancer is the most common and most frequency cancer in women, and the prevalence rate is increasing stably and it brings the highest mortality in women. The breast cancer not only reduces the quality of women’s life but also a great burden to the global health system. Breast cancer is a kind of disease involved a complex and multiple processes. Though epidemiological studies have provided risk factors of breast cancer(such as inherited mutations, hormones, environment factor and so on). The traditional therapy of breast cancer is surgery, radio therapy, chemotherapy, and with the development of molecular biology, target therapy has become a novel treatment in breast cancer. But the single target therapy is limited in clinical because of the drug-tolerance and side-effect. So it is a hot spot to develop a proper combination therapy, as we can increase efficiency of chemotherapeutics drugs and improve quality of life at the same time. Ellagic acid, a natural phenolic phytochemicals, extracted from raspberries, strawberries, walnuts, pomegranates, grapes and so on. Previous work showed properties of pomegranate extracts in cancer therapy and ellagic acid(EA) is the part and the metabolic product of pomegranate extracts, this make EA could be a potential chemoreceptive agent in cancer research as well. Experiments in vivo and vitro have evidenced that EA could induce apoptosis and cell cycle arrest in cancer cells, inhibit angiogenesis, inflammation, lipid peroxidation, liver fibrosis, and radical scavenging activity. But there is no clear mechanism about its effect of anti-neoplasm. As the relationship of PI3 K and breast cancer is gradually established, the PI3 K pathway act as a therapeutic target is increasingly matured. The small molecular PI3 K inhibitor GDC-0941 is in a phase of clinical trial. In this study, we are exploring the synergistic effect between ellagic acid and PI3 K inhibitor(GDC-0941) on breast cancer cell lines including proliferation, apoptosis induction, migration and possible mechanisms.Methods:(1) Cell proliferation assay and Colony formation assay evaluate EA combined GDC-0941 effect on inhibiting growth of breast cancer cells.(2) Apoptosis assay to test the combination effect of EA and GDC-0941 with a panel of breast cancer cell lines.(3) characteristic protein of apoptosis, c-PARP, is detected by western blot.(4)p-S6 RP and 4EBP1, which is the down-stream of PI3K-m TOR signaling pathway is checked by western blotting assay.(5) Scratching assay is used to test the migration inhibiting of the combination.(6) To exam the efficacy of EA in combination with GDC-0941 against mouse breast cancer cell line(D419) transfer to lung in vivo.Results:(1) EA and GDC-0941 combination is efficacious in a panel of breast cancer cell lines on proliferation and growth.(2) EA combined with GDC-0941 promoted apoptosis of breast cancer cell lines and brings on an augment of cleaved PARP.(3) EA and GDC-0941 combined treatment disrupted PI3 K and m TOR downstream 4EBP1 and S6 RP signaling effectors.(4)EA shows a synergistic effect with GDC-0941 in the migration inhibiting of cells.(5)EA and GDC-0941 combined therapy suppressed mouse breast cancer cell line(D419) transfer to lungs in vivo.