核酸适配子探针对转移性大肠癌细胞的多靶点成像及其靶标的定量分析

发布时间：2018-06-24 01:34

  本文选题：核酸适配子 + 转移性大肠癌　； 参考：《高等学校化学学报》2016年07期

【摘要】：利用核酸适配子对肿瘤细胞的高亲和力靶向识别功能以及量子点的高荧光发射强度和光稳定性等特性,制备了识别不同靶点的核酸适配子探针,将其联合使用实现了对肿瘤细胞的多靶点成像及其靶标的定量分析.使用通过Cell-SELEX技术筛选得到的可特异性识别转移性大肠癌细胞系Lo Vo的7个核酸适配子,分别与量子点QD605偶联制备分子探针.基于流式细胞术的竞争实验结果表明,7个探针可特异性识别靶细胞的不同靶点,相互之间无识别干扰.对靶细胞的荧光成像表明,与单一探针相比,多个探针联合使用可明显提高细胞表面的荧光信号强度,且阳性细胞检出率明显增多,显示出更高的检测灵敏度.使用流式细胞术及荧光成像定量方法分析了7个探针对不同转移特性大肠癌细胞系的识别能力,结果表明,多个探针联合使用可有效评价大肠癌细胞的转移潜能.本研究证实通过多个核酸适配子探针的联合使用可有效提高对靶细胞识别的灵敏度和准确性,为核酸适配子的广泛应用及大肠癌的靶向诊断提供了新的思路和手段.
[Abstract]:Based on the high affinity target recognition function of aptamers to tumor cells and the high fluorescence emission intensity and photostability of quantum dots, aptamer probes with different targets were prepared. The multitarget imaging of tumor cells and the quantitative analysis of their targets are realized by using the combined use of the two methods. Seven aptamers of metastatic colorectal cancer cell line Lo Vo were screened by Cell-SELEX technique and coupled with QD605 to prepare molecular probes. The results of competitive experiments based on flow cytometry show that the seven probes can specifically identify the different targets of the target cells, and there is no interference between them. The fluorescence imaging of target cells shows that compared with a single probe, the combined use of multiple probes can significantly improve the fluorescence signal intensity on the cell surface, and the positive cell detection rate is obviously increased, showing a higher detection sensitivity. The recognition ability of 7 colorectal cancer cell lines with different metastatic characteristics was analyzed by flow cytometry and quantitative fluorescence imaging. The results showed that the combined use of multiple probes could effectively evaluate the metastatic potential of colorectal cancer cells. This study proved that the combination of multiple aptamer probes can effectively improve the sensitivity and accuracy of target cell identification, and provide a new idea and means for the wide application of nucleic acid aptamers and the targeted diagnosis of colorectal cancer.
【作者单位】： 中国医科大学卫生部细胞生物学重点实验室医学细胞生物学教育部重点实验室细胞生物学教研室;
【基金】：国家自然科学基金(批准号:21375149,81502703) 沈阳市科技局资助项目(批准号:F13-220-9-29)资助~~
【分类号】：O657.3;R735.34
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本文编号：2059379