Tanaka-Okamoto Miki, Mukai Mikio, Takahashi Hidenori, Fujiwara Yoshiyuki, Ohue Masayuki, Miyamoto Yasuhide
Department of Molecular Biology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.
Department of Multiphase Health Screening, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan.
Glycobiology. 2017 May 1;27(5):400-415. doi: 10.1093/glycob/cww133.
Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. The candidates comprised 9 core1 and 5 core2 glycans. All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. To fully evaluate the clinical utility of these candidates, a further verification study is required.
进行了针对硫酸化O-聚糖的糖组学分析，以鉴定新型血清碳水化合物肿瘤标志物。通过对血清中制备的吡啶氨基化聚糖进行α-神经氨酸酶消化，然后进行阴离子交换色谱，富集硫酸化聚糖。通过两种高效液相色谱分离构建硫酸化O-聚糖谱。将20名健康对照者的谱与11名胃癌患者和9名胰腺癌患者的谱进行比较，确定了14个候选标志物。使用包括酶消化和质谱在内的各种方法对这些候选物的结构进行了精确分析。这些候选物包括9种核心1聚糖和5种核心2聚糖。所有这些候选物均为单硫酸化，其中11种还进行了单岩藻糖基化或双岩藻糖基化，并且包含各种决定簇，如6-磺酸化2型乳糖胺、6-磺酸化Lewis X、6-磺酸化Lewis Y、3'-磺酸化1型乳糖胺和3'-磺酸化Lewis A。此外，在核心1聚糖中，有5个候选物显示出1型和2型乳糖胺杂合主链。使用选择反应监测测定法定量了所有40名受试者血清中这些候选聚糖的水平。这些分析表明：（i）所有候选物的水平在至少一名或多名患者的血清中升高；（ii）具有6-磺酸化Lewis X、6-磺酸化2型乳糖胺或3'-磺酸化Lewis A的1型-2型杂合主链的核心1候选物在多种患者的血清中升高；（iii）候选物的水平在患者之间差异很大，这表明分析多个候选物将是筛查各种癌症的有效手段。为了全面评估这些候选物的临床实用性，需要进一步的验证研究。
