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p53基因对人肺癌细胞系耐药性的影响
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p53基因对人肺癌细胞系耐药性的影响
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p53基因对人肺癌细胞系耐药性的影响
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Journal Article
p53基因对人肺癌细胞系耐药性的影响
2008
Overview
背景与目的肺癌细胞的耐药性是影响化疗疗效的重要因素之一,已有的研究表明p53基因异常与肿瘤细胞耐药性的产生密切相关。本研究的目的是探讨不同状态的p53基因对人肺癌细胞系耐药性的影响,为临床选择有针对性的化疗药物提供依据。方法采用质粒pEGFP,分别构建含正义、反义p53cDNA的真核细胞表达载体,测序法检测外源p53基因。用脂质体介导转染人肺癌细胞系801D,建立含不同p53基因状态的单克隆细胞系,分别为pEGFP-801D、pEGFP-sensep53-801D[含有正义p53,pEGFP-p53（RS）-801D]、pEGFP-antisensep53-801D[含有反义p53,pEGFP-p53（AS）-801D]。荧光显微镜观察细胞绿色荧光,PCR法检测外源基因,MTT法检测耐药性,流式细胞术检测细胞周期与凋亡。结果测序结果显示外源p53基因分别以正向、反向连接于载体,荧光显微镜下转染细胞可见绿色荧光,PCR法检测出外源p53基因和neo基因。pEGFP-p53（AS）-801D细胞系对顺铂（Cisplatin,DDP）的IC50为0.26±0.09μg/mL,显著低于母系（0.55±0.19μg/mL,P〈0.05）和空载系（0.77±0.13μg/mL,P〈0.05）。pEGFP-p53（RS）-801D细胞系对DDP的IC50为0.43±0.25μg/mL,显著低于空载（P=0.000）但高于pEGFP-p53（AS）-801D（P〈0.05）。pEGFP-p53（RS）-801D细胞系对紫杉醇（Paclitaxel,TAX）的IC50为2.34±0.43ng/mL,显著低于母系（8.40±1.50ng/mL,P﹤0.05）,低于空载系但无显著差异。pEGFP-p53（RS）-801D细胞系对5氟尿嘧啶（5-Fluorouracil,5FU）的IC50为2.08±0.18μg/mL,显著低于母系（4.90±1.12μg/mL,P〈0.05）和空载系（3.41±0.86μg/mL,P〈0.05）。DDP能诱导pEGFP-p53（AS）-801D出现G2期阻滞,TAX、5FU诱导pEGFP-p53（RS）-801D细胞系分别出现G2、S期阻滞,DDP、TAX能诱导细胞凋亡和死亡。结论肺癌细胞中不同状态的p53基因对化疗药的耐药性有不同影响。p53突变、缺失与DDP耐受相关,p53缺失与TAX、5FU耐受
