Research On The Apoptosic Action On Nude Mice With The High Metastatic Human HCC Transplanted In Situ (LCI-D20)of Ginsenoside Rg3Alone And When Combined With Oxaliplatin And Sorafenib

Objective:To observe the effects of the apoptosic actionon nude mice with the high metastatic human HCC transplanted in situ (LCI-D20) of Ginsenoside Rg3alone and When Combined With Oxaliplatin And Sorafenib, and to explore the expression of apoptosis related proteins,, to provide the theoretical foundation for the combination of chemotherapy with traditional medicine and antiangiogenic therapy to cure hepatocarcinomaMethods:Using the nude mice with the high metastatic human HCC transplanted in situ(LCI-D20) established by the HCC institute of the Fudan University,60in-situ bearing nude mice were divided into8groups which each contains7-8. Group A:Ginsenoside Rg3alone (5mg/kg,2w),Group B:oxaliplatin alone (5mg/kg,q2d,2w),Group C:Sorafenib (30ml/kg,2w), D:Ginsenoside Rg3(5mg/kg,2w)combine with Sorafenib (30ml/kg,2w); E:Ginsenoside Rg3(5mg/kg,2w)combine with oxaliplatin (5mg/kg, q2d,2w),F:Sorafenib (30ml/kg,2w) combine with oxaliplatin (5mg/kg, q2d,2w); G:Ginsenoside Rg3(5mg/kg,2w)combine with Sorafenib (30ml/kg,2w) and oxaliplatin(5mg/kg, q2d,2w);the control group,Group F: NS(10ml/kg,2w). Experiment started on the11th day after the tumor transplantation.All groups were treated with abdominal injection per day for2weeks, then killed and dissected, inhibitory effect on the transplantation tumor and use a variety of experimental methods to observe its effect on the transplanted tumor cells:(1) using Inverted microscope to observe the cell morphology changes;(2)Test each transplanted tumor cell apoptosis rate through Tunel assay;(3)Using Immunohistochemical SP Staining to obver the expression of Caspase-3, Survivin,Bax,Bcl-2;(4) Using Westing Blotting to obver the expression of Caspase-3, Survivin,Bax,Bcl-2.Results:(1) Effect on inhibiting tumor growth:A,B,C,D,E,F and G groups inhibited the transplanted tumor significantly. The inhibitory percentage of each group were20.60%,31.68%,34.71%,48.64%,52.08%,36.97%,51.95%,and According to weeb coefficient, Ginsenoside Rg3combine with oxaliplatin and Sorafenib has synergism effect,oxaliplatin and Sorafenib has addition effect,three drugs has addition effect;(2)USMarsdenia tenacissima injection combine with oxaliplatin and endostar has synergism effect.Although the three drugs groups compare with two drugs groups had synergism tendency but no marked difference between each other.(2) Using TUNEL to calculate the AI:that compared with NS group, each group has the obvious effect of transplanted tumor cells apoptosis, drug induced apoptosis in three effect is stronger (P<0.01). Compared with the single drug group, the two drugs combination group were significantly induce apoptosis (P<0.01); Compared with single medicine group and two drug groups each, three drug combination group significantly induce apoptosis (P<0.01, P<0.05).(3)Immunohistochemical results showed that the treatment group can significantly reduce the Bcl2protein expression, compared with saline control group, the difference was statistically significant (P<0.05); But compared with saline control group, Caspase3, the expression of Bax and Survivin difference is not obvious.(4) Western bloting) experiments show that:compared with normal saline group, each group of Caspase3and Bax protein expression and Bax/Bcl-2ratio have increased (P<0.05), Survivin and Bcl2protein expression is decreased significantly (P<0.05).Conclusion:(1) the ginseng saponins Rg3on nude mouse transplantation tumor of liver cancer have certain inhibition of liver cancer cell growth, and promote the role of cell apoptosis, but relatively weak, and oxaliplatin into its tumor suppression effect is obvious enhancement after combination, show the synergies effect; In ginseng saponins Rg3, on the basis of oxaliplatin into use after sorafenib, showed the increase trend.(2) the ginseng and the mechanism of ginsenoside Rg3treatment effect of liver cancer can be through the Caspase3, Bax apoptosis related gene expression, by Survivin, Bcl-2expression, as well as raising Bax/Bcl-2ratio and implementation, but the specific mechanism remains to be further in-depth study.(3) This research provided the theory and experimental foundation for the combination of TCM with western medicine to cure hepatocarcinoma.