Effects Of Antiangiogenesis Of Sorafenib In Combination With Thalidomide On Proliferation Of Hepatocarcinoma Cell And Chick Chorioallantoic Membrane

Background : Hepatocellular Carcinoma( HCC) is a common malignant tumor diseases in our country, due to large number of hepatitis b,hepatitis c infection in our country and poor sanitation in some parts, the inc idence of liver cancer in China accounts for about half of the world.Because of its pathogenesis is hidden, in about 90% of cases have been found miss operation time. Statistics show that, even in patients with surgical removal, the recurrence rate in 5 years is up to 61.5% at most. It is in urgent for medical workers to look for a better treatment for patients with advanced liver cancer. Malignant tumor growth is rapid. Its growth is dependent on the nutrition provided by tumor vascular. Targeted drug resistance of liver cancer cell proliferation and tumor angiogenesis becomes hot research topic in today’s non-operative therapy of liver cancer. Sorafenib is a new targeted therapy drugs, developed by the German bayer,which has been FDA approved for system of clinical treatment of liver cancer.. Sorafenib has a dual antitumor : 1,Directly inhibit the proliferation of tumor cells through blocking the Ras/Raf/MEK/ERK signaling pathway. 2,Iindirectly inhibit the growth of tumor cells by blocking tumor angiogenesis through reducing VEGF tyrosine kinase receptor(VEGFR) activity.Thalidomide, which initially worked as sedatives controlling pregnant women during pregnancy, is disabled due to serious birth defects events. But in recent years, the study found that thalidomide has the characteristics of anti-angiogenesis and inhibiting tumor, which have applied in the treatment of solid tumor. In the treatment of tumor, the effect of the drug alone is usually limited. In order to achieve better treatment effect and less side effects, more and more studies focus on the combination. Such as Sorafenib combined 5 – FU or doxorubicin, Thalidomide combined paclitaxel in the treatment of advanced liver cancer, etc. But the Combination of Sorafenib In and Thalidomide in the treatment of liver cancer research is rarely seen in the report.Objective: To study the effects of antiangiogenesis of sorafenib in combination with thalidomide on chick chorioallantoic membrane(CAM) and the proliferation inhibition of of sorafenib in combination with thalidomide on BEL 7402 cell. We hope to confirm that the joint application of sorafenib and thalidomide in terms of anti-tumor growth and anti-angiogenesis has more advantages.Providing theoretical basis for further clinical test and treatment.Methods:1,The influence of combination on hepatocellular carcinoma cell proliferation. With BEL7402 cell as experimental object, conducting sorafenib into different concentration of working fluid. Through the MTT method figure out the inhibition rate and the IC50 value of sorafenib. We work out the IC50 value of thalidomide in the same way. Then using the logarithmic phase of BEL7402 r cells to fulfil 96- well plates with 200 ul per hole and 0.2×105cells/ml. There are four groups:control group, sorafenib, thalidomide, combination. Each group have 5 holes and a total of five 96- well plates. After 24 hours hatching,dosing respectively in the cases of incubation. Random once every 12 hours, taking out 1 piece of 96- well plates, and were determined by MTT experiment. Figure out the A value of each group with the instrument, and then calculate the cell growth inhibition rate in each group. Finally drawing the growth curve of five days and evaluate the effect of the combination. 2,The effects of antiangiogenesis of sorafenib in combination with thalidomide on chick chorioallantoic membrane(CAM). Eggs incubated for 7 days were made into CAM models. Divided the models into four groups: control group,sorafenib,thalidomide, sorafenib with thalidomide. After dosing process,each model was incubated for 2 days continuely. The specimens were fixed,took pictures,and the vascular coverage of each specimen was counted. And analys ising the advantages of combination evaluation after statistics processing.Results :1,The IC50 value of sorafenib and thalidomide.After treating BEL7402 cell for 48 hours with 1.5, 3,6, 12, 24,μΜ/L sorafenib the inhibition rate were12.5%、22.2%、34.9%、49.2%、65.1%. After treating BEL7402 cell for 48 hours with 6.25, 12.5, 25, 50, 100 μg/ml thalidomide, the inhibition rate were18.2%、29.1%、38.2%、58.2%、69.1%. The IC50 value of sorafenib and thalidomide are 11.9μΜ/L and 36.9μg/ml.2, The influence of combination on BEL7402. The influence of combination is greater than the s ingle drug group. The differences are statistically significant(p < 0.05). 3,The influence of combination on Chick Chorioallantoic Membrane. The microscopic observations show that: the thick dense, more branches.Blood vessels can be seen in control group, while in dosing group,there was obviously less than control group. And blood vessels have few branches. Part of the vascular contraction fracture, which is the most obvious in combination. Though the analys is of image software,we found that the comparison difference between s ingle drug groupand control group was statistically significant[(30.2±2.9)vs(38.3±2.7), q=17.05, p<0.05;(26.5±2.1)vs(38.3±2.7), q=24.59, p<0.05]. The comparison difference between combined drug group and single drug group was statistically significant[(12.6%±1.5%)vs( 30.2%±2.9%), q=36.68, p<0.05;(12.6%±1.5%)vs(26.5%±2.1%), q=28.67, p<0.05 ].Conclusion: Either of sorafenib and thalidomide has obvious inhibition of proliferation on BEL7402 cel, and both have obvious concentration-response relationship. But the proliferation inhibitory effect of combination drugs group is more apparent. Either of sorafenib and thalidomide has inhibitive effectof angiogenesis in CAM. But when used in combination with both,it would be far more inhibitive.