| Background: Melanoma is one of the most aggressive skin tumors in the world,characterized by high invasiveness,easy metastasis,poor prognosis and high mortality.Nowadays,there are many treatments for melanoma.According to the different stages of the tumor,different surgical methods are the main methods for the treatment of melanoma,others include radiotherapy,chemotherapy,molecular targeted therapy,immunotherapy,etc.However,these treatments have many side effects and high drug resistance.Therefore,it is of great significance to further study the drugs and methods for the treatment of the tumor,and to look for a new treatment to improve the prognosis of patients is a process of continuous efforts.Extracts from nature,with unique pharmacological effects,a wide range of distribution,relatively low side effects,and low prices,therefore,have a broad application prospects in the treatment of cancer.Fucoidan has a variety of biological activities,including anti-tumor,anti-inflammatory,anti-oxidation,anti-coagulation,anti-viral,anti-angiogenesis and immune modulation.Furthermore,a large number of in vitro experiments and animal experiments showed that fucoidan has antitumor effects on lung cancer,colon cancer,breast cancer,gastric cancer,hepatoma cancer and mouse melanoma cells,however,the effect of fucoidan on human melanoma cells has not been reported.The previous work of our research grouphas concluded that fucoidan can inhibit A375 cell proliferation and promote apoptosis.Objective:On the basis of the previous work,we continue to study the effect of fucoidan on human melanoma A375 cell line,the effects on the cell cycle,and the possible mechanism of action,whether there is the involvement of Bcl-2 / Bax,p-Erk pathway,and the expression of the related protein level,to provide experimental and theoretical basis for the clinical application of fucoidan in the treatment of melanoma.Methods : Respectively at the concentration of 0?g/ml(control group),50?g/ml,100?g/ml,200?g/ml and400 ? g/ml of fucoidan treatment of human melanoma A375 cells after 24 hours,then cell morphology was observed under light microscope,changes of cell cycle were observed with flow cytometry using Annexin V-FITC/PI double labeling method,and changes of Apoptosis protein Bcl-2 / Bax,phosphorylated extracellular regulated protein kinase(p-Erk)and related proteins were detected by Western blot.Results: After treatment of A375 cells with different concentrations of fucoidan 24 h,the cells in G2/M phase increased with the increase of fucoidan concentration;however,there was no significant change in the percentage of S phase and G0/G1 phase.The proportion of G2/M phase of A375 cells in the blank control group(0 ?g/ml)accounted for 5.63 + 0.08%,the G2/M phase of the experimental group(50?g/ml?100?g/ml?200?g/ml ?400?g/ml)was 9.61±0.23%,12.61±1.25%,20.27±1.75%,29.17±2.01%,respectively.Western blot showed that the apoptosis inhibitory protein Bcl-2 and phosphorylation of extracellular regulated protein kinase(p-Erk)protein levelsto increase as drug concentration decreased;the protein level of pro apoptotic protein Bax was opposite.Conclusion:Fucoidan can obviously induce the arrest of human melanoma A375 cells in G2/M phase and inhibit the mitosis of the cells;the drug can inhibit the proliferation of A375 cells,block the cell cycle in G2/M phase,and promote cell apoptosis,one of the possible mechanisms is to reduce the ratio of Bcl-2 / Bax and down regulate the expression of p-Erk. |
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