Graphene Oxide-Facilitated Comprehensive Analysis of Cellular Nucleic Acid Binding Proteins for Lung Cancer
Graphene Oxide-Facilitated Comprehensive Analysis of Cellular Nucleic Acid Binding Proteins for Lung Cancer
复制标题
氧化石墨烯促进肺癌细胞核酸结合蛋白的综合分析。
DOI：
10.1021/acsami.8b05428
复制
发表时间：
2018-05-30
影响因子：
9.5
通讯作者：
Xiao, Hua
中科院分区：
文献类型：
--
作者：
Shang, Zhi;Qian, Liqiang;Xiao, Hua
核酸结合蛋白（NABP）介导广泛的基本细胞功能。然而，由于缺乏高效的方法，全面提取全细胞NABP是非常具有挑战性的。为此，碳纳米材料，包括氧化石墨烯（GO），羧化石墨烯（cG），和羧化碳纳米管（cCNT），在这项研究中，通过一种新的策略来提取细胞NABP。我们的数据表明，GO，cG，cCNT可以提取近100%的细胞DNA在体外。相反，它们的RNA提取效率分别为60%、50%和29%，这部分解释了为什么与cG和cCNT相比，GO具有最高的NABP产率。我们进一步发现，RNA与纳米材料的功能基团之间的阳离子介导的离子键促进了RNA在纳米材料上的吸附。从GO富集的NABP样品中成功鉴定了约2400种蛋白质，与细胞裂解物相比，88%的注释NABP至少富集2倍，表明我们的策略具有高选择性。该方法进一步应用于比较两种具有不同肿瘤进展能力的肺癌细胞系中的NABP。根据无标记定量结果，共发现118个NABP差异表达，并通过免疫分析成功验证了6个候选NABP，包括ACAA2、GTF 2I、Vim、SAMHD 1、LYAR和IGF2BP 1。测量肺癌患者血清中SAMHD 1的水平，其在癌症进展时显著增加。我们的研究结果表明，GO是一种理想的NABP选择性提取的纳米材料，可广泛用于各种生理和病理生理环境。
Nucleic acid binding proteins (NABPs) mediate a broad range of essential cellular functions. However, it is very challenging to comprehensively extract whole cellular NABPs due to the lack of approaches with high efficiency. To this end, carbon nanomaterials, including graphene oxide (GO), carboxylated graphene (cG), and carboxylated carbon nanotube (cCNT), were utilized to extract cellular NABPs in this study through a new strategy. Our data demonstrated that GO, cG, and cCNT could extract nearly 100% cellular DNA in vitro. Conversely, their RNA extraction efficiencies were 60, 50, and 29%, respectively, partially explaining why GO has the highest NABPs yield compared to cG and cCNT. We further found that ionic bond mediated by cations between RNA and functional groups of nanomaterials facilitated RNA absorption on nanomaterials. About 2400 proteins were successfully identified from GO-enriched NABPs sample, and 88% of annotated NABPs were enriched at least 2 times compared to cell lysate, indicating the high selectivity of our strategy. The developed method was further applied to compare the NABPs in two lung cancer cell lines with different tumor progression abilities. According to label-free quantification results, 118 differentially expressed NABPs were discovered and 6 candidate NABPs, including ACAA2, GTF2I, VIM, SAMHD1, LYAR, and IGF2BP1, were successfully validated by immunoassay. The level of SAMHD1 in the serum of lung cancer patients was measured, which significantly increased upon cancer progression. Our results collectively demonstrated that GO is an ideal nanomaterial for NABPs selective extraction, which could be broadly used in varied physiological and pathophysiological settings.