Mechanism Of Proliferation And Apoptosis By Turmeric Volatile Oil On Human Melanoma Cell Line A375

Objective The recent research reports indicate that turmeric volatile oil has inhibitory effects on tumor cells,can significantly reduce the number of tumor,reduce the tumor volume,the main role in the tumor early can prevent tumor formation.In this paper,to study the effect of apoptosis and proliferation by turmeric volatile oil(TVO)on human melanoma cell line A375 cells,and to explore its mechanism.Methods The different concentration of TVO worked on A375 cells in vitro.Cell proliferation assay(CCK8)method to measure the toxicity and proliferation inhibition rate at different concentration(0-160mg/L)and time(24h,48 h,72h);According to the CCK8 results,the experiment was conducted to select the effective minimum drug concentration:Giemsa staining observes the morphology of cell apoptosis under inverted microscope;cell apoptosis detected by DNA fragmentation;cell apoptosis was detected by flow cytometry;Western blot detects Caspase-3 and BIRC7 protein expression.Results TVO has significant inhibitory effects on the growth of A375 cells in a time dose dependent relationship,CCK-8 detection showed that the concentration of TVO was between(0?2.5?5?10?20?40?80?160mg/L),after24 h,the inhibition rates were(0.00±0.00?0.015±0.02?0.084±0.02?0.087±0.01?0.096±0?0.087±0.01?0.122±0.03?0.209±0.01);48h(0.00±0.00?0.005±0.00?0.139±0.02?0.160±0.01?0.217±0.03?0.225±0.02?0.246±0.01?0.355±0.01);72h(0.00±0.00?0.323±0.00?0.446±0.01?0.470±0.02?0.479±0.02?0.352±0.01?0.385±0.02?0.441±0.01),vs 0mg/L,P<0.05.TVO induced apoptosis in A375 cells by morphological observation,drug induced cell prolife-ration after slow,adherence decreased,cell gap increases,shape became irregular,nuclear pyknosis,nuclear fragmentation and karyolysis,extraction of DNA gel electrophoresis drugs for48 h,the typical apoptotic DNA ladder,and with the increase of the concentration ofthe more obvious nuclear fragmentation;flow cytometry to measure the cell apoptosis rate,concentration of 0,2.5,5,10mg/L TVO,A375 apoptosis rate was 12.7,13.6,15.4%,the mortality was 0.1%;Western blot(0,2.5,5,10mg/L)TVO concentration was measured after 48 h,with the increase of drug concentration,the expression of apoptosis related proteins Caspase-3,Caspase-1,TNR-alpha increased gradually,and the expression of apoptosis related proteins BIRC7,TAK1,JNK1,TAB1 decreased gradually,compared with 0mg/L,P<0.05.Conclusion TVO can inhibit the cell proliferation,and promote cell apoptosis on human melanoma cell line A375 cells.One of its mechanisms is through the down-regulation of the protein expression of BIRC7,which induced cell apoptosis and activated Caspase-3 of apoptosis pathway key enzyme,thereby inhibiting tumor cell differentiation,proliferation and promoting apoptosis.