| Background and Objective:Malignant tumors are a serious threat to human health.Primary hepatocellular carcinoma,accounting for 90% of liver cell carcinoma,is one of the highest incidence of malignancy in the world.The mortality rate is the second highest in China.Surgical resection is the most effective treatment,but liver cancer usually found in the late stage,it is difficult to remove by surgery,and the recurrence rate is very high.The current treatment of liver cancer drugs have serious side effects,and the treatment effect is not ideal,which is far from meeting clinical needs.Therefore,the search for new safe and effective antineoplastic drugs has significant social and economic benefits.Apoptosis is a procedural death that is actively controlled by genes.The mitochondria mediate the internal apoptosis pathway and the death receptor-mediated external apoptosis pathway are the two main pathways for the occurrence of apoptosis,both of which eventually lead to apoptosis by caspase-3 activation.Cell apoptosis and inhibition of proliferation of cancer cells are closely linked,and provide a new way to develop more anti-cancer drugs.S-5 is a 3-oxo-1,2-naphthoquinone analogue,which has been shown to have a good antitumor effect.On the basis of primary work,this study explored the effectand mechanism of S-5 on the proliferation and apoptosis of Huh-7 human hepatocellular carcinoma cells,and provided the theoretical basis for the further development of S-5 as anti-hepatocarcinoma drugs.Methods:Effects of different concentrations of S-5(0.94~15 ?mol·L-1)on Huh-7 cells cultured in vitro,MTT assay was used to detect the cell growth inhibition rate.Apoptosis rate was detected by FCM(Annexin V-FITC/PI)double staining method.The apoptotic rate was measured by acridine orange/ethidium bromide(AO/EB)double fluorescence staining and transmission electron microscopy.The effect of S-5on mitochondrial membrane potential was detected by JC-1 method.The expression of Bcl-2,Bax,Caspase-3 and Fas,FasL and TNF-? in Huh-7 cells were detected by Western blot.The expression of S-5 was detected by JC-Protein expression.Results:1.S-5 significantly inhibited the proliferation of Huh-7 cells(P<0.05).The IC50 values of 24 h,48h and 72 h were 3.955?mol·L-1,3.522?mol·L-1,1.659?mol·L-1,showing a significant dose and time-dependent effect.2.The apoptotic rate of Huh-7 cells was significantly increased after S-5treatment(P <0.05),the apoptotic rates of the cells after 48 h treatment with 0.94,1.88,3.75 and 7.5?mol·L-1were(7.37±0.40)%,(34.70±1.32)%,(76.87±1.46)%,(95.0±2.0)% respectively,and showed a dose-and time-dependent effect.3.Apoptotic cells morphological changes occur,the emergence of apoptotic bodies and cytoplasmic fragments.4.S-5 induced a decrease in mitochondrial membrane potential in a dose-dependent manner.The ratio of JC-1 polymer to monomer was 11.64±3.36,5.69±1.17,2.04±1.54 and 0.67±0.24 after treated with 0.94,1.88,3.75 and 7.5?mol·L-1 for 48 h.5.S-5 can inhibit the expression of Bcl-2 protein and up-regulate the expression of Bax,Caspase3,Fas,FasL and TNF-? protein in a dose-dependent manner.Conclusion:1.S-5 significantly inhibited the proliferation of Huh-7 cells in vitro.2.Induced apoptosis is an important mechanism of S-5 inhibition of Huh-7cell proliferation.3.S-5 can induce apoptosis of Huh-7 cells through mitochondrial apoptosis pathway and death receptor pathway. |
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