Establishment Of Human Hepatocellular Carcinoma Multidrug Resistant Cell Line BEL7404/ADM

Objective:To establish multidrug resistant strain of human hepatocellular carcinoma BEL7404 and to study its biological characteristics.Methods:The cell line BEL7404 was induced to form a multidrug resistant cell subline(BEL7404/ADM)by exposing it to gradually increase concentration of ADM.MTT assay was used to evaluate drug sensitivity and to measure cell growth.Flow cytometric assay was employed to examine cell cycle and to determine drug influx and efflux.Results:The resistant index of BEL7404/ADM is as much 68.7 times as that of BEL7404.The doubling time of BEL7404/ADM was 6.49 hours longer than BEL7404.The assay of Rhodamine 123 expressed that the function of P-gp in BEL7404/ADM was higher than that of its drug-sensitive strain.Conclusions:BEL7404/ADM was human multidrug-resistant cell line and its biological characteristics had been changed greatly.Its multidrug resistance may be mainly correlated with a higher expression of P-gp. Objective:To investigate the effects of the combination of transfected wild type p53(wt-p53)gene and Adriamycin(ADM)on drug resistance in human hepatocellular carcinoma cell line.Methods:The cells were divided into four groups:the control group,the wt-p53 group(the p53 transfected group),the ADM group(the ADM-treament group),the associated group(p53 transfected with ADM-treatment group).The MTT assay was used to analyse the growth of BEL7404 and BEL7404/ADM cells.The apoptosis of these cells were evaluated by flow cytometry(FCM).Results:BEL7404 cells and BEL7404/ADM cells were inhibited 52.53% and 47.01%respectively with wt-p53 gene transfected on the fourth day.At the same time,in the associated group,BEL7404 cells and BEL7404/ADM cells were inhibited 73.86%and 69.15%respectively.By the assay of FCM,in BEL7404 strain,an increased G₁ phase proportion was induced in the associated group.The percentage of cell apoptosis in the associated group was highest in four groups.In BEL7404/ADM strain,G₁ phase arrest was observed in the associated group and its apoptosis percentage was highest.There was no significant difference in the growth and apoptosis between the two cell lines of the associated group.Conclusions:The growth of hepatocellular carcinoma drug-resistant strain and drug-sensitive strain were obviously inhibited in the group of wt-p53 gene and ADM.And the associated group benefited the apoptosis.But there was no significant difference in the growth and apoptosis between the two cell lines of p53 transfected and ADM-treatment group.