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目的:探讨染料木素(genistein)对非小细胞肺癌A549/DDP细胞的耐药逆转及对肺耐药相关蛋白(LRP)和多药耐药相关蛋白(MRP)表达的影响。方法:采用细胞计数法绘制A549/DDP及A549细胞生长曲线;MTT法测定12.5μg/ml染料木素预处理对A549/DDP细胞耐药的逆转作用,并在荧光显微镜下观察细胞形态变化;用RT-PCR和Western blotting分别检测染料木素对肿瘤细胞LRP和MRP的mRNA及蛋白表达的影响。结果:A549/DDP和A549细胞生长曲线相似,倍增时间分别为(27.38±0.25)h和(18.15±0.36)h;染料木素预处理后耐药倍数从2.28倍下降至1.57倍;镜下观察细胞形态出现体积过大、伪足过度伸展、细胞边缘模糊等变化;LRP及MRP mRNA表达均出现明显下调(P<0.05),但二者蛋白表达无明显改变。结论:A549/DDP细胞基本保留了亲代细胞的生长特性;染料木素可以逆转A549/DDP细胞的耐药性,作用机制可能与下调LRP及MRP mRNA表达有关。
Objective: To investigate the reversal of drug resistance of genistein to non-small cell lung cancer A549 / DDP cells and its effect on expression of lung resistance-related protein (LRP) and multidrug resistance-related protein (MRP). Methods: The growth curve of A549 / DDP and A549 cells was drawn by cell counting method. The reversal effect of 12.5μg / ml genistein pretreatment on A549 / DDP cells was determined by MTT assay. The morphological changes of cells were observed under fluorescence microscope. The effects of genistein on mRNA and protein expression of LRP and MRP in tumor cells were detected by RT-PCR and Western blotting respectively. Results: The growth curves of A549 / DDP and A549 cells were similar with the doubling time of (27.38 ± 0.25) h and (18.15 ± 0.36) h, respectively. The genotoxicity of genistein decreased from 2.28 to 1.57 fold after pretreatment with genistein. The morphological changes of cell morphology such as overabundance of hyperextension, excessive extension of pseudopodia and blurring of cell edge were observed. The expressions of LRP and MRP mRNA were significantly down-regulated (P <0.05), but the expression of LRP and MRP did not change significantly. CONCLUSION: The growth characteristics of parental cells are basically preserved in A549 / DDP cells. Genistein can reverse the drug resistance of A549 / DDP cells. The mechanism may be related to the down-regulation of LRP and MRP mRNA expression.