Anti-tumor Effect And Pro-apoptotic Molecular Mechanism To Tumor Cells Of Danggui Beimu Kushen Decoction Combined With Cis-platin

Objective: To study the separate and combined tumor inhibition effect of DangguiBeimu Kushen decoction (DBKD) and Cis-diammin-odichloroplatinum(DDP) in treatingH22hepatocellular carcinoma xenograft KM mice, investigate apoptotic molecularmechanism of DBKD to oncocyte in treating tumor-bearing mice through the detection ofsome apoptosis-associated molecular expression levels and serum enzyme, in order toprovide scientific experimental basis for anti-tumor treatment clinically with DBKD.Methods: Sccessful KM mice H22hepatocellular carcinoma xenograft models wererandomized into Model Control Group (MCG), DDP control group (DDPCG), DBKDgroup (DBKDG), DBKD plus DDP group (DBKD-DDPG). There was also a normalcontrol group (NCG). During the15d treatment, observe the general state of the mice,obtain the bodyweight of the mice, long and short diameters of the tumor. The mice weregiven water and not given food from18:00on the15thd of treatment. On the next day after15d treatment, blood, tumor, liver, kidney and thymus were obtained from sacrificed micefor blood routin, tumor inhibition rate, q value, serum biochemical test for LDH, AST, ALT,AKP and BUN, CR, ELISA detection of tumor tissue HIF-1α, pathological detection oftumor tissue, liver, kidney, thymus, immunohistochemistry detection of tumor tissue p53and caspase3protein and qRT-PCR detection of bax, bcl-2mRNA.Results:①From the gross observation, DBKDG was better than MCG,DBKD-DDPG was better than DDPCG. Male mice tumor inhibition rates of DBKDG,DDPCG, DBKD-DDPG were34.1%,63.4%,75.6%respectively, q≈1.00; female micetumor inhibition rates of DBKDG, DDPCG, DBKD-DDPG were33.3%,73.3%,80.0%, q≈1.01②Compared with MCG, the level of tumor tissue HIF-1α, mutant p53protein(p<0.01)and bcl-2mRNA in DBKDG was significantly lower; the level of caspase3protein(p<0.01)and bax mRNA in DBKDG were significantly higher.③Compared withDDPCG, the indexes of thymus, white cell number, expression of tumor tissue caspase3protein (p<0.01)and bax mRNA in DBKD-DDPG were significantly higher; the level oftumor tissue HIF-1α and the activity of serum LDH, AST, ALT, AKP and BUNconcentration in DBKD-DDPG were significantly lower (p<0.01), CR was lowed but not statisticly different（p>0.05）; pathologic injury to hepatic lobule, renal tubule andthymocyte were lesser in DBKD-DDPG; the expression of tumor tissue mutant p53proteinand bcl-2mRNA were not lowered significantly，but bax/bcl-2ratio was increasedsignificantly.Conclusion:①DBKD has anti-proliferation effect to H22hepatocellular carcinomaxenograft, which might be related to down regulation of HIF-1α, mutant p53protein andbcl-2mRNA, up-regulation of caspase3protein and bax mRNA.②Combinedapplication of DBKD and DDP can synergisticly enhance anti-proliferation effect to H22hepatocellular carcinoma xenograft of DDP, which might be related to the up-regulation ofcaspase3protein and bax/bcl-2ratio, down-regulation of tumor tissue HIF-1α.③Combined application of DBKD and DDP can reduce DDP-induced damage to organs andtissues, which might be related to the protection of normal tissue cell membrane leading toless release of LDH, AST, ALT, AKP into blood and up-regulation of white cell number.