Résumé
Objective To investigate the effect of Ganlong capsule on axillary graft tumour in Balb/c nude mice.Methods BEL-7402,a human hepatocellular carcinoma sensitive strain,was inoculated into the armpit of nude mice for modeling.After successful modeling,mice were divided into model group,sorafenib group,CII-3 group,degreasing cream group,and Ganlong capsule high-dose,medium-dose and low-dose groups,and normal nude mice were included in blank group.All groups were given continuous administration for 14 days.The tumor inhibition rate,organ index,biochemical index,anoxic microenvironment related proteins and their mRNA expression levels were compared among all groups.Results All drug treatment groups could inhibit tumor growth,and the average tumor inhibition rate was the highest in Ganlong capsule medium-dose group.Compared with the model group,the heart index,kidney index,liver index and lung index of nude mice in all drug treatment groups were increased,and the spleen index were decreased,but most of the changes were little.Various biochemical indexes showed that Ganlong capsule was safe and did not cause obvious tissue damage.The mRNA expressions of hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR2)in sorafenib group and Ganlong capsule medium-dose group were significantly lower than those in model group(P<0.05).Immunohistochemical results showed that all drug treatment groups showed different degrees of inhibition on VEGFR2,HIF-1α,VEGF,Ki-67 and CD31 protein expressions,among which sorafenib group,Ganlong capsule medium-dose group and CII-3 group had stronger inhibitory effects.Conclusion Ganlong capsule can effectively inhibit the growth of axillary graft tumour in Balb/c nude mice,and down-regulate the expression level of hypoxic microenvironment related protein and their mRNA in the graft tumor.
Résumé
OBJECTIVE：To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3（shorted for “degreasing cream ”and“CⅡ-3”）reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS ：MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib （positive control ），degreasing cream and C Ⅱ-3 on HepG2/ADM cells ，then IC 20 was calculated. The experiment was divided into sensitivity drug ，drug-resistance group ，sorafenib group，degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ，and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ，and other 3 groups were treated with relevant medicine （IC20 as drug concentration ）. The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp （expression produce of MDR1 gene），LRP，BCRP] and that related to enzyme-mediated multidrug resistance pathway （GST-π and Topo Ⅱ）. RESULTS ：The IC 20 of degreasing cream ，CⅡ-3 and sorafenib were （2.40±0.16）， （200.44±27.52），（18.00±1.82）μg/mL，respectively. Compared with sensitivity group ，the protein expressions of Bcl- 2，P-gp， LRP，BCRP and Topo Ⅱ，the mRNA expressions of MDR 1， LRP，BCRP and GST-π were increased significantly in drug resistance group （P＜0.05 or P＜0.01）. Compared with @qq.com drug-resistance group ，the mRNA and protein expression of MDR1 mRNA and LRP ，BCRP，GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group（P＜ 0.05 or P＜0.01）；the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased （P＜0.01）， while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group （P＜0.05）. CONCLUSIONS：Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux ， promoting cell apoptosis ，reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.