Tanaka-Okamoto Miki, Hanzawa Ken, Mukai Mikio, Takahashi Hidenori, Ohue Masayuki, Miyamoto Yasuhide
Department of Molecular Biology, Osaka International Cancer Institute, 3-1-69, Otemae, Chuo-ku, Osaka 541-8567, Japan.
Department of Medical Checkup, Osaka International Cancer Institute, 3-1-69, Otemae, Chuo-ku, Osaka 541-8567, Japan.
Anal Biochem. 2018 Mar 1;544:42-48. doi: 10.1016/j.ab.2017.12.026. Epub 2017 Dec 19.
We previously identified four glycan tumor marker candidates using a HPLC-based method. One candidate was sialyl Tn (STN), NeuAcα2-6-GalNAc. In this study, glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine. Pyridylaminated-STN levels of 147 gastric cancer, 85 pancreatic cancer and 10 cholangiocarcinoma patients together with 102 normal controls were accurately quantified using HPLC separation followed by selected reaction monitoring (SRM) assay, which used a stable isotope, tetradeuterium-labeled pyridylamino glycan as an internal standard. Additionally, STN values were also quantified using conventional competitive inhibition radioimmunoassay (RIA). The two STN levels determined by RIA and SRM gave a similar distribution pattern in sera. STN levels were increased in sera from cancer patients compared to those from normal controls. Moreover, the STN levels in sera of cancer patients determined by the two different assay procedures showed a good correlation (i.e., correlation coefficient >0.9). Our results suggest it may be better to determine STN levels using SRM instead of RIA.
我们之前使用基于高效液相色谱的方法鉴定出了四种聚糖肿瘤标志物候选物。其中一个候选物是唾液酸化Tn(STN),即NeuAcα2-6-GalNAc。在本研究中,通过肼处理从血清中制备聚糖,随后用氨基吡啶进行荧光标记。使用高效液相色谱分离,然后采用选择反应监测(SRM)测定法,以稳定同位素四氘代吡啶氨基聚糖作为内标,对147例胃癌患者、85例胰腺癌患者、10例胆管癌患者以及102例正常对照者的吡啶氨基化STN水平进行了准确量化。此外,还使用传统的竞争性抑制放射免疫测定法(RIA)对STN值进行了量化。通过RIA和SRM测定的两种STN水平在血清中呈现出相似的分布模式。与正常对照者相比,癌症患者血清中的STN水平升高。此外,通过两种不同测定方法测定的癌症患者血清中的STN水平显示出良好的相关性(即相关系数>0.9)。我们的结果表明,使用SRM而非RIA来测定STN水平可能更好。