金纳米荧光分子信标对活细胞中STAT5b mRNA的检测  

Gold nanoparticle-based fluorescent molecular beacon for the detection of STAT5b mRNA expression in living cell

作　　者：李洋[1] 薛建鹏[1] 崔思思[1] 田俊梅[1] 章冬银[1] 曹洁[1] 顾月清[1] 

机构地区：[1]中国药科大学生命科学与技术学院生物医学工程教研室,南京210009

出　　处：《科学通报》2013年第7期537-544,共8页Chinese Science Bulletin

基　　金：国家自然科学基金(30970776;81000666;31050110123)资助

摘　　要：信号转导和转录激活因子5b(signal transducer and activator of transcription5b,STAT5b)是JAK-STAT途径中重要的一种转录因子.该转录因子与肿瘤的发生发展密切相关,正日益受到人们的关注.在本研究中,优化STAT5b mRNA序列并合成出一种新型STAT5b金纳米荧光分子信标.该信标由发卡型STAT5b mRNA特异性序列、FITC荧光染料、金纳米粒三部分组成.通过激光共聚焦和流式细胞术等实验方法,实现该信标对HepG-2细胞和PC12细胞中STAT5b mRNA的实时检测.最后,运用RT-PCR方法,该分子信标检测STAT5b mRNA的可靠性得到验证.基于本研究实验结果,可推断该新型金纳米荧光分子信标有望进行JAK-STAT信号通路中其他转录因子的检测.Signal transducer and activator of transcription 5b （STAT5b） is an important protein in JAK-STAT signal pathway and is responsible for the metastasis and proliferation of tumor cells. In this study, we demonstrated a novel beacon which consisted of a gold nanoparticle core functionalized with a dense monolayer of hairpin DNA with a specific sequence for the STAT5b mRNA and modified with FITC. By the confocal laser scanning microscope and flow cytometry, we successfully detected the expression of STAT5b mRNA using this novel beacon in HepG-2 cells and PC12 cells. At last, to confirm the presence and absence of STAT5b mRNA in HepG-2 cells and PC12 cells, RT-PCR applification was performed. This strategy is a promising approach for the intracellular measurement of transcription factors, RNA or proteins.

关 键 词：金纳米粒 分子信标 STAT5B MRNA 

分 类 号：R450[医药卫生—治疗学]