| Objective:The aim of this study is to firstly establish a nude mice transplantedtumor model of Oral squamous carcinoma multidrug resistance cell(KB/ADM),and.then detect the expressions of multidrug resistance(MDR1)and'Topoisomerase ?,?a,??(TOPO ?,?a,??)gene and significance intransplanted tumor cells.Methods1.With different concentrations of ADM,VCR,VP-16 and HCPT treatment KB and KD/ADM cells,resistance determined by MTT assay to determine its value-added inhibition rate of tumor cells and IC50 and multiple drug resistance.2.With adriamycin(ADM)for induction,by gradually increasing dosage,establish multidrug resistance KB/ADM cell lines.1×1010/L,0.2mL KB/ADM cells were subcutaneously transplanted into the nude mice's right neck.The models of KB/ADM in nude mice were randomly group and received NS,VP-16(5mg/kg),NC(5mg/kg),VP-16(5mg/kg)+VRP(5?mol/L),NC(5mg/kg)+VP-16(5mg/kg)of desired drug and dosage for 10 days.The reversal effect of nitidine chloride was evaluated by tumor weight and volume.3.Morphology of xenografts were investigated by HE staining.4.The apopotosis of tumor organization was detected by TUNEL assay.5.The expressions of MDR1,Topo ?,Topo ??,Topo ?? mRNA levels were detected by real-time PCR.Results1.With different concentrations of ADM,VCR,VP-16 and HCPT treatment KB and KD/ADM cells,the IC50 value of KB cells were 65.77?g/mL,46.04?g/mL,38.73?g/mL and 47.78?g/mL;the IC50 value of KD/ADM cells were 159.91p,g/mL,87.27?g/mL,78.46?g/mL,129.43?g/mL.The four kinds of anticancer drugs to deal with KB cells was obviously higher than these of KB/ADM cells;Relative to the KB cell,KB/ADM multiples drug resistence were 2.43,1.90,2.03,2.71.2.Prior to the nude mice were sacrificed,none in both groups was dead.Before delivery,the growth speed,tumors of nude mice had no difference between the volume(P>0.05).But after the treatment had obviously difference(P<0.05).3.1 'Compared with negative control group(NS),VP-16 group,the strucutal of squamous cell carcinoma KB/ADM transplanted tumor in Nude Mice of the group of NC,VP-16+VRP,NC+VP-16 had obvious changed by HE assay.3.2 Effect of different treating group of NS,VP-16,NC,VP-16+VRP,NC+VP-16 apoptosis index(AI)respectively were 0.00%,5.45%,16.60%,48.03%,60.13%.4.After different treatment,the mRNA expressions variation of gene MDR1,Topo ?,Topo ??,Topo ?? of squamous cell carcinoma KB/ADM transplanted tumor in Nude Micewere were down-regulated.Conclusion:Transplanted tumor model of KB/ADM drug resistance cell in nudemice is successfully established.NC can obviously enhance etoposide induce drug-resistant cell apoptosis,and inhibit the mRNA expression gene MDR1,Topo ?,Topo?a,Topo ??.NC may reverse multi-drug resistance of tumor,make oral squamous carcinoma resistant cells KB/ADM increased sensitivity of VP-16. |
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