本文選題：核酸適配子 + 轉(zhuǎn)移性大腸癌�。� 參考：《高等學(xué)�；瘜W(xué)學(xué)報》2016年07期

【摘要】：利用核酸適配子對腫瘤細(xì)胞的高親和力靶向識別功能以及量子點的高熒光發(fā)射強度和光穩(wěn)定性等特性,制備了識別不同靶點的核酸適配子探針,將其聯(lián)合使用實現(xiàn)了對腫瘤細(xì)胞的多靶點成像及其靶標(biāo)的定量分析.使用通過Cell-SELEX技術(shù)篩選得到的可特異性識別轉(zhuǎn)移性大腸癌細(xì)胞系Lo Vo的7個核酸適配子,分別與量子點QD605偶聯(lián)制備分子探針.基于流式細(xì)胞術(shù)的競爭實驗結(jié)果表明,7個探針可特異性識別靶細(xì)胞的不同靶點,相互之間無識別干擾.對靶細(xì)胞的熒光成像表明,與單一探針相比,多個探針聯(lián)合使用可明顯提高細(xì)胞表面的熒光信號強度,且陽性細(xì)胞檢出率明顯增多,顯示出更高的檢測靈敏度.使用流式細(xì)胞術(shù)及熒光成像定量方法分析了7個探針對不同轉(zhuǎn)移特性大腸癌細(xì)胞系的識別能力,結(jié)果表明,多個探針聯(lián)合使用可有效評價大腸癌細(xì)胞的轉(zhuǎn)移潛能.本研究證實通過多個核酸適配子探針的聯(lián)合使用可有效提高對靶細(xì)胞識別的靈敏度和準(zhǔn)確性,為核酸適配子的廣泛應(yīng)用及大腸癌的靶向診斷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.
【作者單位】： 中國醫(yī)科大學(xué)衛(wèi)生部細(xì)胞生物學(xué)重點實驗室醫(yī)學(xué)細(xì)胞生物學(xué)教育部重點實驗室細(xì)胞生物學(xué)教研室;
【基金】：國家自然科學(xué)基金(批準(zhǔn)號:21375149,81502703) 沈陽市科技局資助項目(批準(zhǔn)號:F13-220-9-29)資助~~
【分類號】：O657.3;R735.34
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本文編號：2059379