The Research Of Aidi Injection Reverses Multidrug Resistance Of Human Hepatocellular Carcinoma

Objective:Explore whether Aidi injection reverses multidrug resistance(MDR)of hepatocellular carcinoma Bel7402/5-FU cell line.And explore potential mechanisms.Methods:1.Establish the multidrug resistance human hepatocellular carcinoma Bel-7402/5-FU by using increasing concentrations of fluorouracil(5-FU).2.Divide the experiment into two groups when the resistant cell 7402/5-FU successfully established.It is Bel7402 cell and 7402/5-FU cell.The two group of cells grown in ninety-six well plates respectively.And then divide each cells into seven groups after the cells sidewall stability.Each cell of the first group join normal culture medium.The remaining six groups of cells join the culture medium which containing Fluorouracil and Doxorubicin and Mitomycin and Methotrexate and cyclophosphamideand and cis-Dichlorodiamineplatinum.Each hole volume for 200 ul.After twenty-four hours.Multiscan Spectrum reads the absorbance values at the wavelength of 450 nm.Calculate the half maximal inhibitory concentration(IC50)of every chemotherapy drugs by using the Statistical Package for Social Science nineteen(SPSS 19.0).Calculate seven chemotherapy drugs inhibition ratio(IR)to Bel-7402 and Bel-742/5-FU and the drug resistance index(RI)of Bel-7402/5-FU.3.Through the different concentration of AIDI injection acting on the Bel-742/5-FU cell.The method of cck-8 detect whether the AIDI injection has inhibitory effect on the Bel-742/5-FU cell.4.Divide the experiment into three groups.It is Bel-7402 cell and Bel-742/5-FU cell and the Bel-742/5-FU cell which was effected by AIDI injection.Extract the total protein in three groups.Using the western blot method calculate the expression of the protein levels of multi-resistance related 1(MRP1)p-glycoprotein(P-gp)and programmed cell death 5(PDCD5)in Bel-7402 and Bel-742/5-FU and handled by half lethal concentration of Aidi injection.Results:1.The half lethal concentration of Bel-742/5-FU Significantly increased than Bel-7402 to six chemotherapy drugs,showed different levels of drug resistance to six drugs,2.The aidi injection has a certain inhibitory effect to Bel-7402/5-FU cell line.And the inhibitory rate increasing with the stronger concentration.3.The protein expression levels of MRP1 and P-gp in Bel-7402/5-FU cells were obviously higher than those in Bel7402 cells.while the expression of PDCD5 protein in Bel7402/5-FU cells was lower than that in Bel7402 cells.Through half maximal inhibitory concentration the aidi injection acting on the Bel-7402 and Bel-7402/5-FU cell line,compared with the Bel7402 cell lines.The Bel-7402/5-FU cell line which handled by the the half maximal inhibitory concentration of AIDI injection.The expression of MRP1 and P-gp proteins was down-regulated after Aidi injection treatment,while the PDCD5 protein was significantly incrased.the difference has statistically significance(P<0.05).Conclusion:The Aidi injection can reverse multi-drug resistance of the Bel-7402/5-FU cell line.The mechanisms associated with lower the expression of MRP1 and P-gp and promote the expression of PDCD5.