Effect Of Proteasome Inhibitor MG132 With Cisplatin On The Xenografts On The Nude Mice Of Hela Cells And On The Expression Of The P65 In Hela Cells

Objective:To observe the effect of proteasome inhibitor MG132 combined with cisplatin on the xenografts of Hela cells on the nude mice and to test the NF-κB's expression of Hela cells.Methods:1 Inoculability in nude mice:twenty female nude mice were fed in the animal center.Cells were cultured to logarithm phrase and digested with EDTA and then adjusted density of cells to 4×10~7/ml.We inoculated the cells on the right anterior limbs of the nude mice by hypodermic injection under the sterile condition.In the tenth day,we divided the nude mice into four stochastic groups.There were 5 mice in every group and managed by the way there in after.(1)control group:0.2ml 0.9%saline, i.p,(2)MG132 group:MG132 5mg/kg,0.2ml,i.p,(3)cisplatin group:cisplatin 3mg/kg,0.2ml i.p,(4)cisplatin with MG132 group:MG132 5mg/kg,0.2ml and cisplatin 3mg/kg,0.2ml i.p.2 Inspection of xenograft's growth,volume and weight.Inspection of pathology and histology:some of xenografts were fixed by 10% formaldehyde.Then they were dehydrated and embed.After slicing up and HE staining,they were observed by light microscope.3 Western blotting was used to detect the expression of p65 in HeLa cells:HeLa cells were divided into four groups:the first group was not treated by cisplatin or MG132,it was just cultured by culture medium;the second group was treated by 5μmol/l MG132 for 48h;the third group was treated by 2.5μg/ml cisplatin for 48h;the forth group was treated by 5μmol/l MG132 combined with 2.5μg/ml cisplatin for 48h.Results:1 When the nude mice were sacrificed,the volume of the xenografts of every group are:control group:1004±239.83mm~3,MG132 group:863±192.52mm~3,cisplatin group:551±143.60mm~3,the MG132 with cisplatin group were 236±71.64mm~3(P＜0.001).the weight of the xenografts of every group are:control group:0.86±0.12g,MG132 group:0.74±0.13g,cisplatin group:0.49±0.08g,the MG132 with cisplatin group:0.27±0.04g.The analysis of variance was used to examine the differences between the four groups on the volume and the weight of the xenografts on the nude mice.The differences reached the statistical significance.(p＜0.05)2 Comparing with the control group,the tumor inhibiting rates of MG132 group,cisplatin group and the MG132 with cisplatin group were 15%,48%,81%,the inhibiting rates of the weight of the xenografts were 9%,43%,69%respectively.3 Compared with the control group,the p65's expression of he cisplatin group obvious up-regulated(P＜0.05).Compared with the control group,the p65's expression of the MG132 and the MG132 with cisplatin groups obvious down-regulated(P＜0.05).But the differences between the MG132 and the MG132 with cisplatin groups did not reach the statistical significance.(p＞0.05).Conclusion:1 MG132 can inhibit the xenografts of Hela cells on the nude mice and can increase the inhibition effect of cisplatin to them.2 Cisplatin can increase the expression of p65 in Hela cells,while Proteasome inhibitor MG132 can decrease the expression of p65,which may have relationship with the phenomenon that MG132 increase the inhibition effect of cisplatin to Hela cells.