DNA aptamers from whole-serum SELEX as new diagnostic agents against gastric cancer.
DNA aptamers from whole-serum SELEX as new diagnostic agents against gastric cancer.
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全血清 SELEX 中的 DNA 适体作为胃癌的新诊断剂
DOI：
10.1039/c8ra08642g
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发表时间：
2019-01-02
期刊：
影响因子：
3.9
通讯作者：
Huang, Baihai
中科院分区：
文献类型：
--
作者：
Zheng, Yue;Zhao, Yunwang;Di, Ya;Xiu, Chenlin;He, Lei;Liao, Shiqi;Li, Dongdong;Huang, Baihai
胃癌仍然是全球癌症死亡的主要原因之一。尽管诊断方法有所改进，但迄今尚未实现早期发现。胃癌的早期诊断可显著提高患者的治愈率。因此，需要一种新的诊断方法。本研究分别以胃癌患者血清和正常人血清为靶血清和阴性血清，采用消减SELEX技术筛选胃癌血清特异性DNA适体。采用全血清消减SELEX技术，构建了4个胃癌血清特异性核酸适体Seq-3、Seq-6、Seq-19和Seq-54，其Kd值分别为128 ± 26.3 nM、149 ± 23.6 nM、232 ± 44.2 nM和202 ± 25.6 nM。这些产生的适体通过区分正常血清但与其他癌症血清密切相关而对其靶血清显示出更高的特异性。将筛选出的4种高亲和力DNA适体进一步应用于建立基于qPCR的胃癌早期检测方法。此外，我们进行了MALDI-TOF MS，然后进行二级肽测序MS分析，以鉴定适体结合蛋白。在这些潜在的生物标志物中，APOA 1、APOA 4、PARD 3、Importin亚基α-1显示出相对较高的评分概率。因此，本研究所制备的四种ssDNA适体有望成为胃癌诊断的分子探针。
Gastric cancer is still among the leading causes of cancer deaths worldwide. Despite the improvements in diagnostic methods, the status of early detection has not been achieved so far. Early diagnosis of gastric cancer may significantly improve the cure rate of patients. Therefore, a new diagnostic method is needed. In this study, subtractive SELEX was performed to screen gastric cancer serum-specific DNA aptamers by using gastric cancer serum and normal serum as the target and negative serum, respectively. Four highly specific aptamers generated for gastric cancer serum, Seq-3, Seq-6, Seq-19 and Seq-54, were developed using whole-serum subtractive SELEX technology with Kd of 128 ± 26.3 nM, 149 ± 23.6 nM, 232 ± 44.2 nM, 202 ± 25.6 nM, respectively. These generated aptamers showed higher specificities toward their target serum by differentiating normal serum but closely related other cancer serums. The selected four high affinity DNA aptamers were further applied to the development based on qPCR method for the early detection of gastric cancer. In addition, we performed MALDI-TOF MS followed by secondary peptide sequencing MS analysis for the identification of the aptamer binding proteins. Among these potential biomarkers, APOA1, APOA4, PARD3, Importin subunit alpha-1 showed a relatively high score probability. Therefore, these four ssDNA aptamers generated in our study could be a promising molecular probe for gastric cancer diagnosis.
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