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采用多重实时聚合酶链反应检测RRM1、XRCC1、TUBB3和TS mRNA以预测非小细胞肺癌对放化疗的反应

Multiplex real-time PCR for RRM1, XRCC1, TUBB3 and TS mRNA for prediction of response of non-small cell lung cancer to chemoradiotherapy.

作者信息

Wu Guo-Qiu, Liu Nan-Nan, Xue Xiu-Lei, Cai Li-Ting, Zhang Chen, Qu Qing-Rong, Yan Xue-Jiao

机构信息

Center of Clinical Laboratory Medicine of Zhongda Hospital; Institute of Biotechnology and Clinical Pharmacy, Southeast University, Nanjing, China E-mail :

出版信息

Asian Pac J Cancer Prev. 2014;15(10):4153-8. doi: 10.7314/apjcp.2014.15.10.4153.

Abstract

BACKGROUND

This study was aimed to establish a novel method to simultaneously detect expression of four genes, ribonucleotide reductase subunit M1(RRM1), X-ray repair cross-complementing gene 1 (XRCC1), thymidylate synthase (TS) and class III β-tubulin (TUBB3), and to assess their application in the clinic for prediction of response of non-small cell lung cancer (NSCLC) to chemoradiotherapy.

MATERIALS AND METHODS

We have designed four gene molecular beacon (MB) probes for multiplex quantitative real-time polymerase chain reactions to examine RRM1, XRCC1, TUBB3 and TS mRNA expression in paraffin-embedded specimens from 50 patients with advanced or metastatic carcinomas. Twenty one NSCLC patients receiving cisplatin- based first-line treatment were analyzed.

RESULTS

These molecular beacon probes could specially bind to their target genes in homogeneous solutions. Patients with low RRM1 and XRCC1 mRNA levels were found to have apparently higher response rates to chemoradiotherapy compared with those with high levels of RRM1 and XRCC1 expression (p<0.05). The TS gene expression level was not significantly associated with chemotherapy response (p>0.05).

CONCLUSIONS

A method of simultaneously detecting four molecular markers was successfully established and applied for evaluation of chemoradiotherapy response. It may be a useful tool in personalized cancer therapy.

摘要

背景

本研究旨在建立一种同时检测四个基因,即核糖核苷酸还原酶亚基M1(RRM1)、X射线修复交叉互补基因1(XRCC1)、胸苷酸合成酶(TS)和III类β微管蛋白(TUBB3)表达的新方法,并评估其在临床中预测非小细胞肺癌(NSCLC)放化疗反应的应用价值。

材料与方法

我们设计了四种基因分子信标(MB)探针用于多重定量实时聚合酶链反应,以检测50例晚期或转移性癌患者石蜡包埋标本中RRM1、XRCC1、TUBB3和TS mRNA的表达。对21例接受以顺铂为基础的一线治疗的NSCLC患者进行了分析。

结果

这些分子信标探针能在均相溶液中特异性结合其靶基因。与RRM1和XRCC1表达水平高的患者相比,RRM1和XRCC1 mRNA水平低的患者对放化疗的反应率明显更高(p<0.05)。TS基因表达水平与化疗反应无显著相关性(p>0.05)。

结论

成功建立了一种同时检测四个分子标志物的方法,并将其应用于放化疗反应的评估。它可能是个性化癌症治疗中的一种有用工具。

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