目的 本研究通過(guò)去甲斑蝥素（NCTD）作用于肺癌順鉑（DDP）耐藥細(xì)胞A549/DDP，探討其對(duì)DDP耐藥的逆轉(zhuǎn)作用及其相關(guān)機(jī)制。方法 應(yīng)用CCK-8法檢測(cè)NCTD對(duì)A549和A549/DDP細(xì)胞活力的影響、DDP對(duì)A549細(xì)胞和A549/DDP的半數(shù)抑制濃度（IC₅₀）、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.

R965

山東省醫(yī)學(xué)科學(xué)院院級(jí)科技計(jì)劃（2016-41）