O-glycosylated clusterin as a sensitive marker for diagnosing early stages of prostate cancer.
O-glycosylated clusterin as a sensitive marker for diagnosing early stages of prostate cancer.
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O-糖基化簇蛋白作为诊断早期前列腺癌的敏感标记。
DOI：
10.1002/pros.24094
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Takashi Ueno
中科院分区：
文献类型：
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作者：
Saiko Kazuno;Tsutomu Fujimura;Makoto Fujime;Yoshiki Miura;Takashi Ueno
前列腺特异性抗原（PSA）是前列腺癌最常用的诊断标志物。然而，在高度可疑的前列腺癌患者中经常出现低PSA值（<10 ng/ml），这削弱了临床检查的准确性。本研究的目的是开发一种更好的诊断前列腺癌的解决方案，以克服PSA的缺点。MethodsWe专注于患者血清蛋白的糖基化状态，并进行了全面的凝集素微阵列分析，以characterzeN ‐ andO‐聚糖使用前列腺癌和良性前列腺疾病的血清。接下来，我们使用凝集素固定化微珠检索具有特征性聚糖结构的候选血清蛋白，并使用称为相对和绝对定量（iTRAQ）标记的同量异位素标签技术通过定量质谱法对其进行鉴定。最后，我们构建了一个新的检测方法来定量一个候选糖蛋白与新鉴定的聚糖。ResultsLectin微阵列分析显示，从前列腺癌患者的血清具有较高的亲和力Jacalin，苋尾（ACA）凝集素，andMaclura pomifera（MPA）凝集素，与良性前列腺疾病患者和正常人相比，提示O-糖基化蛋白在前列腺癌患者血清中更丰富。然后，分离优先吸附在Jacalin-琼脂糖上的血清糖蛋白以及生物素-ACA/和生物素-MPA/链霉亲和素固定化磁珠，用iTRAQ标记，并使用定量质谱法鉴定。结果发现，ACA和MPA可识别的簇蛋白在前列腺癌患者血清中比良性前列腺疾病患者血清中更富集。根据这一发现，我们构建了基于Luminex的测定来定量O-糖基化丛生蛋白，其中首先在抗丛生蛋白抗体固定的珠上捕获总血清丛生蛋白，然后通过生物素-MPA和链霉亲和素-藻红蛋白对测定丛生蛋白相关O-聚糖。当PSA值小于10 ng/ml时，血清MPA识别的clusterin水平有助于区分前列腺癌和良性前列腺疾病。结论PSA值小于10 ng/ml时，血清O-糖基化clusterin水平可作为前列腺癌恶性程度的辅助指标。
BackgroundProstate‐specific antigen (PSA) has been the most popular diagnostic marker for prostate cancer. The frequent occurrence of low PSA values (<10 ng/ml) in patients with highly suspicious prostate cancer, however, has undermined the accuracy of clinical examinations. The aim of this study was to develop a better resolution for diagnosing prostate cancer to overcome the disadvantage of PSA.MethodsWe focused on the glycosylation status of patients' serum proteins and conducted comprehensive lectin microarray analyses to characterizeN‐ andO‐glycans using sera from prostate cancer and benign prostatic diseases. Next, we retrieved candidate serum proteins with characteristic glycan structures using lectin‐immobilized beads and identified them by quantitative mass spectrometry using a technique referred to as isobaric tag for relative and absolute quantitation (iTRAQ) labeling. Finally, we constructed a new assay to quantify a candidate glycoprotein with the newly identified glycans.ResultsLectin microarray analyses revealed that sera from patients with prostate cancer had a higher affinity for Jacalin,Amaranthus caudatus(ACA) lectin, andMaclura pomifera(MPA) lectin, compared with that from patients with benign prostatic diseases and normal subjects, suggesting thatO‐glycosylated proteins are more abundant in sera from patients with prostate cancer. Then, serum glycoproteins preferentially adsorbed onto Jacalin‐Agarose as well as biotin‐ACA/and biotin‐MPA/streptavidin‐immobilized magnetic beads were isolated, labeled with iTRAQ, and identified using quantitative mass spectrometry. It was found that the ACA‐ and MPA‐recognizable clusterin was more enriched in patients' sera from prostate cancer compared with those from benign prostatic diseases. Following this discovery, we constructed a Luminex‐based assay to quantifyO‐glycosylated clusterin, in which total serum clusterin was first captured on anti‐clusterin antibody‐immobilized beads, and then clusterin‐associatedO‐glycans were determined by the pair of biotin‐MPA and streptavidin‐phycoerythrin. When PSA values registered less than 10 ng/ml, the corresponding serum level of MPA‐recognized clusterin determined by this assay was beneficial for distinguishing the patients with prostate cancer from the patients with benign prostatic disease.ConclusionFor PSA values that measure less than 10 ng/ml, the serumO‐glycosylated clusterin level can be a complementary indicator for the malignancy of prostate cancer.