50)、NCTD聯(lián)合不同濃度DDP對(duì)A549/DDP細(xì)胞DDP耐藥性的影響;細(xì)胞克隆形成實(shí)驗(yàn)檢測(cè)NCTD、DDP單給藥及NCTD與DDP聯(lián)合用藥對(duì)A549/DDP細(xì)胞克隆形成率的影響;Western blotting法檢測(cè)A549/DDP和A549細(xì)胞中c-met表達(dá),以及NCTD、DDP單給藥及NCTD與DDP聯(lián)合用藥對(duì)A549/DDP細(xì)胞內(nèi)c-met蛋白表達(dá)的影響。結(jié)果 NCTD劑量相關(guān)性地抑制A549和A549/DDP細(xì)胞的活力,選擇抑制率小于10%的最大NCTD濃度即10 μmol/L作為逆轉(zhuǎn)濃度;DDP作用于A549/DDP和A549細(xì)胞的IC50分別為15.11和5.04 μmol/L,差異有統(tǒng)計(jì)學(xué)意義(P<0.01);A549/DDP細(xì)胞中c-met表達(dá)明顯高于A549細(xì)胞;用NCTD(10 μmol/L)處理后,DDP對(duì)A549/DDP細(xì)胞的IC50從15.11 μmol/L減少到8.06 μmol/L,差異顯著(P<0.01);與DDP單給藥組比較,NCTD聯(lián)合DDP作用明顯降低A549/DDP細(xì)胞克隆形成率(P<0.01);Westernblotting結(jié)果顯示,NCTD聯(lián)合DDP作用明顯抑制順鉑耐藥細(xì)胞A549/DDP細(xì)胞中c-met蛋白的表達(dá),單用DDP或NCTD與對(duì)照組比較差異不明顯。結(jié)論 NCTD與DDP聯(lián)用可逆轉(zhuǎn)A549/DDP細(xì)胞對(duì)DDP耐藥,其機(jī)制可能與抑制c-met蛋白表達(dá)有關(guān)。;Objective To investigate the anti-resistance effects of norcantharidin (NCTD) on cisplatin (DDP) resistance A549/DDP cells and its reversal mechanism. Methods A549 and A549/DDP cells were used as a model for investigating the function of NCTD and DDP in this study.CCK-8 method was used to detect the effect of NCTD on the activity of A549 and A549/DDP cells, IC50 of DDP on A549 and A549/DDP cells, and the effect of NCTD combined with DDP of different concentrations on DDP resistance of A549/DDP cells. Cell clonogenesis assay was used to detect the clonogenesis rate of A549/DDP cells treated with NCTD, DDP alone and the combination of NCTD and DDP; Western blotting was used to detect the expression of c-Met in A549/DDP and A549 cells, and the effect of NCTD, DDP alone and the combination of NCTD and DDP on the expression of c-met protein in A549/DDP cells. Results The cell activity of the A549 and A549/DDP cells were significantly inhibited by NCTD in a dose dependent manner. The concentration of 10 μmol/L was selected to be the reversal concentration, because its maximum rate was less than 10%. The IC50 of DDP on A549/DDP and A549 cells was 15.11 and 5.04 μmol/L respectively, the difference was statistically significant (P < 0.01); The expression of c-Met in A549/DDP cells was significantly higher than that in A549 cells; After treatment with NCTD (10 μmol/ L), the IC50 of DDP on A549/DDP cells decreased from 15.11 to 8.06 μmol/L, the difference was significant (P < 0.01); Compared with DDP alone group, the effect of NCTD combined with DDP significantly reduced the formation of A549/DDP cell clone (P < 0.01). Western blotting results showed that the combination of NCTD and DDP significantly inhibited the expression of c-met protein in cisplatin resistant A549/DDP cells. Conclusion NCTD compared with DDP can reverse the DDPresistance of A549/DDP cells, and its potential mechanism may be ralated to the down-regulation of c-met expression."/>