野黄芩苷通过下调c-met逆转肺癌A549/DDP细胞顺铂耐药

Scutellarin Reverses Cisplatin Resistance in Lung Cancer A549/DDP Cells by Down-regulation of c-met Expression

目的探讨野黄芩苷对肺癌A549/顺铂(cisplatin,DDP)细胞DDP耐药的逆转作用及机制。方法MTT法检测肺癌DDP耐药细胞A549/DDP的耐药倍数、野黄芩苷对其细胞毒性及逆转倍数;平板克隆形成试验检测野黄芩苷对A549/DDP细胞克隆形成的影响;Western blotting检测野黄芩苷对A549/DDP细胞内c-met蛋白表达的影响。结果MTT法检测结果显示,肺癌DDP耐药细胞A549/DDP的耐药性是其亲本细胞A549的5.77倍,差异有统计学意义(P<0.01)。同时,与A549细胞相比,DDP耐药细胞A549/DDP中c-met的蛋白表达显著增高(P<0.01)。野黄芩苷浓度分别为20,40,80和160μmol·L^(-1)时,对A549/DDP细胞的抑制率分别为2.38%,4.01%,7.43%和26.43%,而且随着野黄芩苷药物浓度的增加,克隆形成数目显著减少。选择抑制率<10%的最大野黄芩苷浓度即80μmol·L^(-1)作为逆转浓度;野黄芩苷联合DDP作用可显著降低A549/DDP细胞的耐药性;Western blotting结果显示,野黄芩苷联合DDP作用能显著抑制A549/DDP细胞中c-met蛋白的表达。结论野黄芩苷与DDP联用可提高肺癌A549/DDP细胞对DDP的敏感性,抑制克隆形成,其机制可能与下调c-met蛋白表达有关。

OBJECTIVE To explore the anti-resistance effects of scutellarin on cisplatin(DDP)resistance lung cancer A549/DDP cells and its reversal mechanism.METHODS MTT method was used to detect the reversing multiple of DDP-resistant lung cancer cell line A549/DDP,and cell toxicity,drug-resistant multiple of scutellarin on it;the colony formation assay was performed to determine the effect of scutellarin on the formation of A549/DDP cell clones;Western blotting was used to detect the expression of c-met protein expression in A549/DDP cells treated with scutellarin.RESULTS The result of MTT test showed that the resistance of DDP-resistance cell line A549/DDP was 5.77 times higher than in parent cell A549,the differences were statistically significant(P<0.01).Meanwhile,the protein expression level of c-met was higher in A549/DDP cell line than in A549(P<0.01).When scutellarin concentration was 20,40,80 and 160μmol·L^(-1),its inhibition rates on DDP-resistant cell line A549/DDP were 2.38%,4.01%,7.43%and 26.43%.In addition,the colony formation density was significantly inhibited with the increase of scutellarin concentration.The concentration of 80μmol·L^(-1)was selected to be the reversal concentration,because its maximum rate was<10%;the DDP-resistance significantly decreased in A549/DDP cells after scutellarin and DDP co-treatment.Western blotting results showed that the expression level of c-met significantly decreased in A549/DDP cells after treatment with scutellarin combined with DDP.CONCLUSION Scutellarin combined with DPP can reverse the DDP resistance of A549/DDP cells,inhibit the colony formation and its potential mechanism may be related to the down-regulation of c-met protein expression.