| Background:Hepatocellular carcinoma(HCC) is malignant tumor to severely threan human's healthy.The treatment of liver cancer, especially the advanced liver cancer,Chemotherapy is one of primary therapeutic tool.Sorafenib is,a novel,oral multi-target antineoplastic, to refrain tumor cell proliferation and angiogenesis.It is the first target-drug that can prolong survival time of advanced Hepatocellular carcinoma patients. Owing to the lower efficiency of a single agent, the combined effects of target drug and the traditional chemotherapy drugs become the focus of clinical research. A lot of literature covers that the joint between target drugs and chemotherapy drugs is not a simple superposition, but there exist the most reasonable joint scheme and the best drugs delivery order. Therefore,studying the joint effect mechanism of Sorafenib and the traditional chemotherapy drugs, and looking for ways to further enhance the effectiveness of the best regimen has great practical significance for liver cancer and other solid tumors in the combined therapy.Objective:To research on the different effects that results from the joint administration of Sorafenib and Paclitaxel in different drugs delivery order to the liver cancer cells, thus reveals the mechanism.And to provide theoretical basis for combined therapy of HCC.Methods:Transmission electron microscope observed BEL-7402 to be treated by Sorafenib。The MTT assay to examine the IC50 that Sorafenib effect on the BEL-7402 in 24h,72h and 48h, thus draw the inhibition rate-concentration curve. Flow cytometry to detect different order effect of two drugs which described previously on BEL-7402 cell cycle and apoptosis rate.Western blot analysis to observe Bcl-2 expression level in BEL-7402 cells which treated by the drugs in different administration order.Results:1.Transmission electron microscope observed BEL-7402 cells to be treated with Sorafenib,discovered the cytoplasm shrinkage,chromatin condensation,the formation of apoptotic bodies.2.MTT assay measured IC50 value of Sorafenib to BEL-7402 in 24h, 72h and 48h,were 9.17μg/ml,2.43μg/ml,0.52μg/ ml,respectively.In the same time,with the increasing concentration of drug inhibition rate increase gradually.3.Flow cytometry to show that BEL-7402 cells'G2-M phase was significantly prolonged treated by Paclitaxel alone;treated by Sorafenib S-phase prolonged, G0-G1 phase shortened.The groups of Sorafenib in combination with Paclitaxel in different order showed that all the G0-G1 phase was shortened, Sorafenib was used after group G2-M phase prolonged, S-phase did not change significantly,apoptosis was significantly increased. However,Sorafenib was used before S-phase significantly extended(84.01%),G2-M phase did not change.At the same time joined the two drugs group S and G2-M phase were extended, the apoptosis rate ranged from between the other two groups.4.Western blot analysis discovered the expression of Bcl-2 is obviously decreased in the group which used Sorafenib after Paclitaxel.Conclusions:1.Sorafenib can impact of BEL-7402 cell cycle ,induced the apoptosis of BEL-7402.Reduced Bcl-2 protein expression may be one of the Sorafenib induced apoptosis mechanism.2.Sorafenib can inhibit the BEL-7402 cell line proliferation, and this inhibition was dose and time dependent.3.Sorafenib ahead of Paclitaxel treat BEL-7402 may have lower Apoptosis rate.On the contrary,Paclitaxel ahead of Sorafenib treat can access to higher apoptosis and lower Bcl-2 expression, such an effect may be due to two drugs in the role of the different cell cycle, and the Bcl-2 expression of different, so in clinical treatment of HCC, Paclitaxel in followed by Sorafenib may be given to the better effect.
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