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Relationship between the acquired multi-drug resistance of human large cell lung cancer cell line NCI-H460 by cisplatin selection and p53 mutation
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF TUBERCULOSIS AND RESPIRATORY DISEASES
- Issue:
- 2
- DOI:
- 10.3760/j:issn:1001-0939.2005.02.010
- Key Word:
- 肺肿瘤;药物耐受性;药物疗法;NCI-H460 细胞系;顺铂;基因,p53
Abstract： 目的探讨p53基因突变与肺癌化疗后获得性耐药的关系.方法采用顺铂(DDP)大剂量(50 μmol/L)间歇诱导法,体外诱导具有野生型p53基因的人大细胞肺癌NCI-H460细胞株,建立多药耐药细胞系H460/DDP;用免疫细胞化学法检测P糖蛋白(P-gp)、多药耐药相关蛋白(MRP1)、肺相关蛋白(LRP)、拓扑异构酶Ⅱ(TOPOⅡ)、谷胱甘肽转移酶π(GST-π)及P53蛋白的表达状况;用荧光免疫法测定P53蛋白磷酸化状态;对p53 cDNA全长进行扩增及测序,用含野生型p53基因的质粒pShuttle-CMV-wtp53 cDNA转染耐药细胞,测定转染细胞的药物敏感性.结果新建立的人大细胞肺癌多药耐药株H460/DDP,对DDP及卡铂的耐药指数分别为10.21和9.98,对氟尿嘧啶、多柔比星、表柔比星、甲氨蝶呤、依托泊苷及异长春新碱等有不同程度的交叉耐药.多药耐药株H460/DDP细胞的P53蛋白滞留于细胞浆,在DDP刺激下不能发生磷酸化;LRP表达明显增加(P＜0.05);但其他耐药相关蛋白水平与DDP诱导前相比差异无统计学意义(P＞0.05); 耐药株细胞p53基因第277位点后插入一个"t";耐药株转染pShuttle-CMV-wtp53 cDNA后,其耐药性可发生部分(53.2%)逆转.结论铂类化疗药物诱导的NCI-H460细胞p53基因突变与其对化疗药物耐药性的产生有密切关系,p53基因替代疗法可能是克服这类获得性耐药的一个有效方法.
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