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

关键词:  肺癌  A549/DDP  野黄芩苷  c-met  耐药性

Key words:  lung cancer  A549/DDP  scutellarin  c-met  drug resistance

摘要:

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160 μmol·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80 μmol·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。

Abstract:

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 160 μmol·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 80 μmol·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.