The Compound Traditional Chinese Medicine CFF-1 Induces Prostate Cancer Cell Apoptosis And Autophagy And Its Molecular Mechanism

Prostate cancer(PCa)is the second largest cancers that affect men's health in western countries.According to SEEP(Surveillance,Epidemiology and End Results Program)statistics,there were about 220800 new cases of prostate cancer in the United States in 2015 and 86380 people died of prostate cancer.With the development of economy and the improvement of living standards,the number of people suffering from prostate cancer is increasing year by year.PI3K/AKT is a serine/threonine protein kinase that regulates the survival of the signaling network and affects cellular processes such as cell proliferation,cell survival and gene expression,which makes AKT a potential therapeutic or preventative target for cancer.Over-expression of AKT has been correlated with the development and metastasis of various cancers.Activated AKT disturbs the balance of apoptosis and cell livability by promoting NF-kB and inhibiting the FOXOs pro-apoptotic transcription factor.At present,the domestic and foreign research on the immune regulation of traditional Chinese medicine is a single herb and its monomer components.As a compound Chinese medicine,CFF-1 is less and not enough depth research.In this study,to explore the mechanism of cell death induced by Chinese medicine CFF-1 in vitro and in vivo,the cell viability of prostate cancer cells investigated by morphological observation,MTT assay and CCK-8 assay.respectively.Effect of CFF-1 on cell apoptosis and cell cycle was measured by DAPI staining and Flow Cytometry,respectively.Furthermore,Western blot was employed to investigate the activity of EGFR and its downstream signaling pathways and the levels of the expression of cell apoptosis-related proteins,cell autophagy-related proteins and cell cycle-related proteins.Experiment result showed:(1)Prostate cancer cells and normal cells were treated with different concentrations(0 mg/mL,2 mg/mL,5 mg/mL and 10 mg/mL)of CFF-1,it was found that CFF-1 could inhibit the proliferation and adhesion of prostate cancer cells and the survival rate of cells;(2)The degree of cell apoptosis(including early apoptosis and late apoptosis)was increased with the increasing concentration of CFF-1 and resulted in cell cycle of LNCaP and PC3 cells greatly arrested in G1 phase;(3)At molecular level,CFF-1 competitively targeted EGFR with EGF,and then suppressed auto-phosphorylation of EGFR.Disruption of EGFR activity in turn suppressed downstream PI3K/AKT and RAF-1/MEK/ERK signal pathways and further reduced the level of FOXO1 phosphorylation,finally promoted FOXO1 transcription activity;(4)inhibition of P13K/AKT,RAF-1/MEK/ERK signaling pathway activity and FOXO1 transcriptional activity further inhibited the activity of mTOR/p70S6K signaling pathway and up-regulated the expression of Bim,Fas-L,c-Caspase8,c-Caspase9,c-Caspase3,Bax,c-PARP-1,p21 and p27 while down-regulated the protein levels of cell anti-apoptosis-related proteins Bcl-2,XIAP,Survivin and cell cycle protein Cyclin D1,leading to cell cycle arrest in G1 phase and cell autophagy,finally promoting prostate cancer cells to apoptosis;(5)In vivo experiments further revealed that CFF-1 inhibited the growth of subcutaneous xenografts in nude mice by reducing the protein levels of p-AKT,p-ERK,p-FOXO1 and Cyclin D1 and increasing the protein levels of c-Caspase3,c-PARP-1 and LC3II.In summary,CFF-1 can selectively inhibit the growth of prostate cancer cells,induce autophagy and apoptosis of cancer cells.CFF-1 will be the main candidate drug for clinical treatment of prostate cancer in the future.