| Background and purpose: Primary hepatic caminoma(PHC) is one of the world's most common and most malignant tumor. Currently,chemotherapy,surgery and interventional treatment techniques to liver cancer have been enhanced,but the results are still unsatisfactory.In recent years,Chinese medicine cut a striking figure in the antitumor research with in-depth exploration of new methods of experimental cancer treatment, Based on the theory of traditional Chinese medicine,tumor treatment focus on the overall regulation of the body and playing a comprehensive effect of multiple level,multiple link and multiple targets. It plays an important role whether in suppression and destruction to tumor cells,modulating immune function,improving the signs and symptoms,or reducing the toxicity aspects of chemotherapy and radiotherapy, its potential therapeutic value is incalculable. Therefore,it is very important to place the concrete and effective part and active components of traditional Chinese medicine in the study and to search for new anti-cancer drug which have little toxic side effects but good curative effect in clinical and basic research of liver cancer.Emodin(EM) is one of the active ingredients of rhubarb,giant knotweed rhizome and other Chinese medicine,it showed strong inhibition and killing effects on the liver cancer,breast cancer,lung cancer,prostate cancer and other tumor cells,it also can reverse the multidrug resistance of tumor cells and inhibit tumor angiogenesis and tumor invasion and metastasis. This study was to investigate the effect of emodin on apoptosis and the expression of Cytochrome C(Cyt C),mitochondrial protein Smac/DIABLO(Smac) in vitro in hepatoma carcinoma cells,and further explore the mechanism of tumor cell apoptosis induced by Emodin.Methods:After cultured, human hepatocellular carcinoma SMMC-7721 cell line were randomly divided into 5 groups, intervened by 0,20,40,80μmol·L-1 Emodin respectively and set the positive control group intervened by 100μmol·L-15-fluorouracil (5-Fu).Time of drug effects were all 48 hours. The apoptosis rate was detected by Annexin V-FITC/PI double labeled flow cytometry. Then, the expression of Cyt C and Smac mRNA was measured by Real time quantitative polymerase chain reaction (Real time PCR) and the expression of Cyt C and Smac protein in endochylema was detected by Western blot respectively.Results:Flow cytometry showed that:compared with that of the blank control group, the rate of early apoptotic cells increased markedly,the rate of Living cells was significantly reduced (P< 0.01).And with the increasing concentration of Emodin,early apoptosis rate showed an increasing trend, living cells decreased gradually.At the same time, Real time PCR results showed that:the expression of Cyt C and Smac mRNA significantly increased (P< 0.05). Western blot results show:the protein expression of Cyt C and Smac in endochylema in Emodin groups were higher than that in blank control group(P＜0.01). And with the increasing concentration of emodin, Cyt C,Smac mRNA and cytosolic Cyt C, Smac protein all increased gradually.Conclusions:By promoting synthesis and transcription of Cyt C and Smac and promoting releasing of them from mitochondria to the cytoplasm,Emodin can induce the apoptosis of hepatoma cells through mitochondrial caspase dependent apoptosis pathway to educe anti-tumor effects.
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