目的 建立检测晚期非小细胞肺癌(NSCLC)患者外周血核苷酸切除修复互补基因1(ERCC1 mRNA)表达水平的实时荧光定量逆转录聚合酶链反应(RT-PCR)方法,并探讨该基因表达水平与铂类化疗敏感性的关系.方法 符合入选条件的晚期NSCLC患者60例,化疗前利用实时荧光定量RT-PCR技术对其外周血标本进行ERCC1 mRNA相对定量分析.患者分别接受4～6个周期的含铂类第三代方案的化疗(具体方案为吉西他滨+顺铂或多西他赛+顺铂),并采用实体瘤疗效评价标准(RECIST标准)对患者化疗的有效率(response rate,RR)进行评估,疗效分为完全缓解(CR)、部分缓解(PR)、稳定(SD)和进展(PD).结果 60例患者外周血均检测到ERCC1mRNA的表达.化疗总有效率(PR/PR+SD+PD)30.0％(18/60),总疾病控制率(PR+SD/PR+SD+PD)86.7％(52/60).有效(PR)患者的ERCC1mRNA表达水平(中位值＝1.790)与无效(SD+PD)患者的ERCC1 mRNA表达水平(中位值=2.779)差异有统计学意义(Z=2.376,P＜0.05).疗效为PR,SD,PD患者的ERCC1 mRNA表达水平呈逐渐升高的趋势(中位值分别为1.790,2.883,3.545),差异有统计学意义(H=6.319,P＜0.05).ERCC1 mRNA表达水平低于中位值患者(“低”表达)的有效率(53.3％)与高于中位值患者(“高”表达)的有效率(6.6％)差异有统计学意义(P＜0.05).ERCC1 mRNA表达水平与年龄、性别、ECOG PS评分、临床分期、组织细胞病理学分型等因素之间无显著相关性(P＞0.05).结论 ①外周血细胞ERCC1 mRNA表达水平与铂类化疗敏感性具有显著负相关.②所建实时荧光定量RT-PCR方法能成功检测晚期非小细胞肺癌患者外周血细胞ERCC1 mRNA表达水平,有较高的敏感性与特异性,可应用于临床检测工作,指导临床化疗.%Objective This study tried to develop a method of detecting the expression levels of excision repair cross complement KERCC1) mRNA in peripheral blood by fluorescent real-time RT-PCR, investigate the correlation of the sensitivity of platinum-based chemotherapy in advanced non-small cell lung cancer(NSCLC), provide the basis for predicting clinical outcome from the expression of ERCC1 mRNA. Methods Sixty patients were diagnosed as advanced NSCLC by pathological examination of percutaneous needle mass aspiration or bronchofiberscopy biopsy. The relative quality analysis of ERCC1 mRNA in peripheral blood specimen was performed by the extractive technique of RNA and SYBR Green I fluorescent real-time quantitative RT-PCR assay before chemotherapy. The patients received 4 to 6 cycles of cisplatin-based chemotherapy (GC or DC regimen). The response rate (RR) was evaluated as the chemotherapy finished using Response Evaluation Criteria in Solid Tumors (RECIST) criteria. Results Expression of ERCC1 mRNA was detectable in all patients. The total response rate of cisplatin-based chemotherapy (PR/PR+SD+ PD) was 30. 0%( 18/60) and the total disease control rate (PR+SD/PR+SD+PD) was 86. 7% (52/60); Expression of ERCC1 mRNA in the chemosensitive patients(median level 1. 790) was obviously lower than that in the chemoresistant patients (median level 2. 779)( Z =2. 376, P =0. 017). Expression of ERCC1 mRNA was gradually higher in PR,SD,PD patient groups ( median level 1. 79. 2. 883, 3. 545 , respectively) , there were statistical significant deviation among three groups(H =6. 319, P = 0. 042). There was significant difference in response rate of chemotherapy between the patients with low ERCC1 mRNA expression (RR) and high ERCC1 mRNA expression(RR=53. 3% vs 6. 6% , P 0. 05). Conclusion ①There was negative correlation between ERCC1 mRNA expression in peripheral blood of response rate in advanced NSCLC patients.②The expression of ERCClmRNA in NSCLC was successfully detected by using SYBR Green I fluorescent real-time quantitative RT-PCR assay. This detection method has relatively higher sensitivity and specificity and can be used for clinical analysis.
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