金纳米荧光分子信标在BRCA1 mRNA检测中的应用

Application of Gold-Nanoparticle-Based Fluorescent Molecular Beacons in BRCA1 mRNA Detection

为实时检测活细胞中乳腺癌易感基因1(BRCA1)的信使核糖核酸(mRNA)的表达量,设计并成功制备出一种新型金纳米荧光分子信标。研究了该信标的特异性、灵敏度、稳定性和毒性,并通过激光共聚焦和流式细胞术技术检测了该信标的荧光恢复强度,实现了对人胃癌细胞BGC823和其耐药细胞BGC823/DDP中BRCA1 mRNA表达量的检测。利用逆转录-聚合酶链反应(RT-PCR)技术,验证了该信标用于检测细胞中BRCA1 mRNA表达量的可靠性。结果表明,该信标具有优良特性,能可靠检测出肿瘤细胞中BRCA1 mRNA的表达水平。

In order to detect the expression of messenger ribonucleic acid（mRNA）of the breast cancer susceptibility gene 1（BRCA1））in real time in living cells,a novel gold-nanoparticle-based fluorescent molecular beacon is designed and successfully prepared. The specificity,sensitivity,stability and toxicity of this beacon are investigated,the fluorescence recovery strength of this beacon is detected by the techniques of laser confocal microscopy and flow cytometry,and thus the expression of BRCA1 mRNA in human gastric cancer cell BGC823 and its drug-resistant cell BGC823/DDP is realized.The reverse transcription polymerase chain reaction（RT-PCR）technique is used to verify the reliability of this beacon for detecting the expression of BRCA1 mRNA in cells.The results show that this beacon has excellent characteristics and can be reliably used to detect the expression level of BRCA1 mRNA in tumor cells.