Study On Effect Of Bufalin On The Multidrug-resistant Of Human Bladder Cancer T24 Cells

Objective:To establish a multidrug resistance cell line of human bladder carcinoma(T24/ADM)by intermittent administr ation o f high dose ADM;study the effect of bufalin(BF)on T24/ADM multidrug resistance,and to explore its possible mechanism.Methods:Human bladder carcinoma cell line T24 was induced to multidrug resistance cellline(T24/ADM)by intermittent administr ation o f high dose adriamycin(ADM);Cell morphology was observed by inverted microscope,MTT assay was used to detect the T24/ADM respectively to ADM sensitivity to etoposide(VP-16)and vincristine(VCR),The difference expressions o f mdr-1 gene-coded P-glyeoprotein(Pgp)were examined with real time-PCR(RT-PCR)and flow cytometry(FCM).MTT assay detected the effect of bufalin on the chemotherapeutic sensitivity of T24/ADM cell;FCM detection BF influence on the rate of apoptosis of T24/ADM cells;RT-PCR detection of BF of T24/ADM cell MDR1 effect;FCM detection BF of P-g P expression in T24/ADM cell effect.Results: T24/ADM are resistant to ADM,VP-16,VCR and resistant index were 6.35,3.65,4.24;The expression of MDR1 gene in T24/ADM cells about Parent cells T24(16.28+1.59)times,the rate of positive expression of P-gp of T24/ADM cells was significantly higher than that of the parent cells T24{(17.21+1.53)%VS(0.39+0.13)%},the difference is statistically significant(P<0.05);The resistant index of T24/ADM 10nmol/L after the BF decreased to 2.95,2.04,2.05,the difference is statistically significant(P<0.05),The reversal multiples were 2.75,2.35,2.43;Apoptosis of BF induced by ADM(6.73+1.67)%,and with ADM can significantly increased apoptosis rate(19.94+3.27)%,the difference is statistically significant(P<0.05);The concentration of 10nmol/L,20nmol/L,30nmol/L BF after T24/ADM cells of the MDR1 gene relative expression quantity were 8.32+3.11,4.32+1.26,1.35+0.69,P-g P positive expression rate respectively(11.52+2.29)%,(5.39±1.14)%,(2.69+0.86)%,and the control group without drug of T24/ADM cells compared to(MDR1:16.38+2.58),(P-g P:17.39+3.36)%,the difference has statistical significance(P < 0.05).Conclusion:T24/ADM was a model with multidrug resistance,the T24/ADM cell model of ADM,VP-16 and VCR have resistance.T24/ADM resistant cells of MDR1 gene and P-gp expression was higher than that of T24 cell lines,and the difference is statistically significant(P<0.05).BF induced by T24/ADM resistant cell apoptosis and down regulating the expression of resistance gene MDR1 and resistance protein P-gp,reverse T24/ADM resistant cell multidrug resistance.