Experimental Study On STAT3 Pathway And Other Correlated Genes Of Marsdenia Tenacissima Injection Alone And When Combined With Oxaliplatin And Endostar

Objective:Observe the effects of Marsdenia tenacissima injection alone and the effects when it is combined with oxaliplatin and human recombinant endostatin (endostar) on nude mice with the high metastatic human HCC transplanted in situ (LCI-D20), and investigate the mechanism of STAT3 pathway and someother correlated 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,48 in-situ bearing nu de mice were divided into 6 groups which each contains 8.Group A:Marsdenia tenaciss ima injection alone(15ml/kg,d1-d21),Group B:oxaliplatin alone(5mg/kg,q2d,2w),Group C:Marsdenia tenacissima injection(15ml/kg, d1-d21)combine with oxaliplatin (5mg/kg, q2d,2w), Group D:oxaliplatin(5mg/kg, q2d,2w) combine with human recombinant endost atin (endostar)(10mg/kg,d1-d21),Group E:Marsdenia tenacissima injection(15ml/kg,d1-d21) combine with oxaliplatin(5mg/kg, q2d,2w) and human recombinant endostatin (endostar) (10mg/kg,d1-d21),the control group,Group F:NS(10ml/kg,d1-d21).Experiment started on the 11th day after the tumor transplantation.All groups were treated with abdominal injecti on per day for 3 weeks, then killed and dissected, inhibitory effect on the transplantation tumor and its metastasis to the liver and lung were observed, and the changes of STAT 3,Caspase-3,C-myc,Survivin expression of the transplanted HCC tissue were investigated by using Immunohistochemical SP Staining and real-time PCR.Results:(1)A, B,C, D and E groups inhibited the transplanted tumor significantly. The inhibi-tory percentage of each group were 28.60%,68.91%,73.45%,62.10%,80.14%,and had marked difference with the nagative control group(P<0.05).B,C, D, E groups had marked difference of tumor inhibition with the A group.According to Weeb coefficient,Marsdenia tenacissima injec-tion combine with oxaliplatin has addition effect,Marsdenia 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) Marsde-nia tenacissima injection alone or combined with oxaliplatin and human recombinant endostatin (endostar) could inhibited the invasion and metastasis of the tumor,but there is no marked dif-ference to the negative control group in liver and lung(P>0.05).(3)By immunohistochemical method, the expression of STAT3 and Survivin were down-regulated markedly in each experi-ment group(P<0.05),the expression of Caspase-3 and C-myc were not marked compared with NS group.(4) By real-time PCR, Marsdenia tenacissima injection alone could down-regulated the expression of STAT3,C-myc and Survivin markedly(P<0.05).oxaliplatin alone could down-regulated the expression of STAT3,C-myc and Survivin but not mark-edly(P>0.05).Marsdenia tenacissima injection combined with oxaliplatin could down-regulated the expression of C-myc,up--regulated the expression of Caspase-3 but not markedly(P>0.05). Marsdenia tenacissima injection combine with oxaliplatin and human recombinant endostatin (endostar) could up-regulated Caspase-3 markedly(P<0.05)and down-regulated the expression of C-myc(P>0.05).Conclusion:(1) Marsdenia tenacissima injection could weakly inhibit the proliferation of the high metastatic human HCC transplanted in situ on nude mice, while obviously strengthening effects were observed when combined with oxaliplatin or both with human recombinant en-dostatin (endostar),which indicated that they have synergism tendency.(2) The effects of Mars-denia tenacissima injection which could inhibit the proliferation of hepatocelluar carcinoma may related to down regulation of the expressions of STAT3 and C-myc and Survivin.The effects of inhibiting the proliferation of hepatacellular carcinoma by Marsdenia tenacissima injection alone or combined with oxaliplatin and human recombinant endostatin (endostar) related to STAT3 pathway some degree,such as down regulating of C-myc and up regulating of Caspase-3.The exact mechanism should be invested more until we make it clear.(3)This research provided the theory and experimental foundation for the combination of TCM with western medicine to cure hepatocarcinoma.