Development of New Diagnostic Methods to Recognize the Fucosylated Carbohydrate Chains Expressed by Endometrial Cancer Cells and Clinical Applications

子宫内膜癌的细胞学诊断通常表现为难以解释形态学，因此可能导致假阳性诊断。本研究旨在开发一种新的针对子宫内膜癌特异性表达的碳水化合物抗原（SN-Ag）的酶免疫测定法（EmC-EIA）。子宫内膜标本细胞学标本采用单层标本制备装置“Thin Prep”进行处理。该方法作为子宫内膜癌的辅助诊断方法的有效性在手术和临床标本上进行了评估。手术获得子宫内膜癌44例（G1: 32例，G2: 5例，G3: 5例），正常子宫内膜标本10例。用“Thin Prep”将细胞学标本粘接在滤膜上，浸泡在细胞解离缓冲液中，摇匀后，用超声波对样品进行微分离，制备标本。采用Sandwich EIA法测定样品中SN-Ag的含量，采用特异性检测SN-Ag的单克隆抗体MSN-1。能检测到SN-Ag的样品为阳性样品。72.7% （G1: 75.0%, G2: 80.0%, G3: 40.0%）的子宫内膜癌标本的EmC-EIA阳性，而12例正常子宫内膜标本的EmC-EIA阳性率为0%。在直接观察下获得的手术标本仅包括预期的癌组织，而不包括周围的正常组织或其他细胞类型。这些结果反映为子宫内膜标本的阳性率高于正常子宫内膜标本，正常子宫内膜标本的假阳性率为0%，说明我们的EmC-EIA方法能够准确测量细胞学标本中SN-Ag的含量。子宫内膜标本中EmC-EIA阳性率为69.4% (G1: 7311%, G2: 60.0%, G3: 60.0%)， 137例临床标本中G1阳性率较高，与手术标本阳性率相近。另一方面，84例正常子宫内膜标本的阳性率为15.5%，假阳性率较手术标本有所增加。对这些假阳性病例的仔细重新解释表明，这是由于子宫内膜细胞学标本中包含了宫颈腺细胞。17例子宫腺肌症，阳性率为35.3%；这是在子宫内膜癌和正常子宫内膜标本之间观察到的阳性率。因此，我们的结果表明，我们的EmC-EIA方法能够准确地测量细胞学标本中的SN-Ag量。我们的研究结果还表明，通过对子宫内膜腔进行适当的取样，该方法可以作为辅助诊断方法，并将在临床上适用于评估子宫内膜癌的细胞学标本，特别是高分化的子宫内膜癌，往往表现为难以鉴别诊断。少

Cytologic diagnosis of endometrial cancers often presents with difficult morphologic interpretations and thus may lead to false positive diagnoses. This current study was carried out with the goal to develop a new enzyme immunoassay (EmC-EIA) that targets the carbohydrate antigen (SN-Ag) expressed specifically by endometrial cancers. Cytological specimens obtained from endometrial samplings were processed by the monolayer specimen-preparation apparatus "Thin Prep". The usefulness of this method as an ancillary diagnostic method for endometrial cancer was evaluated on both surgical and clinical specimens. A total of 44 cases of endometrial cancer were obtained at surgery (G1: 32 cases, G2: 5 cases, G3: 5 cases), in addition to10 normal endometrial specimens. Specimens were prepared by adhering the cytological specimens on the filter membrane by "Thin Prep", immersing the specimens in cell dissociation buffer, and after shaking, micro-fractionating the samples by ultrasonographic sonicat … More or. SN-Ag amount in the samples was determined by Sandwich EIA using the monoclonal antibody MSN-1 that specifically detects SN-Ag. Samples in which SN-Ag could be detected were determined to be positive samples. 72.7% (G1: 75.0%, G2: 80.0%, G3: 40.0%) of endometrial cancer specimens were positive by EmC-EIA compared to a 0% positive rate in 12 normal endometrial specimens. Surgical specimens, obtained under direct observation, included only the intended cancer tissue and did not include surrounding normal tissue or other cell types. These results were reflected as a high positive rate among endometrial specimens compared to normal endometrial specimens and as a false positive rate of 0% in the normal endometrial specimens and demonstrated that our EmC-EIA method is able to accurately measure the SN-Ag amount in cytological specimens. Furthermore, a positive rate of 69.4% by EmC-EIA was detected in endometrial specimens (G1: 7311% G2: 60.0%, G3: 60.0%), with a high positive rate detected among G1 cases, from among 137 clinical specimens and was similar to the positive rate's demonstrated with surgical specimens. On the other hand, a positive rate of 15.5% was observed with 84 cases of normal endometrial specimens and showed an increase in the false positive rate compared to surgical specimens. Careful reinterpretation of these false positive cases showed that this was due to inclusion of cervical gland cells in the endometrial cytology specimens. A positive rate of 35.3% was observed with 17 cases of adenomyosis; this is between the positive rate observed for endometrial cancer and normal endometrium specimens. Thus our results demonstrate that our EmC-EIA assay is able to accurately measure SN-Ag amount in cytological specimens. Our results also demonstrate that with proper sampling of the endometrial cavity, this method would serve as an ancillary diagnostic method and would be clinically applicable in the evaluation of cytological specimens for endometrial cancers, especially highly differentiated endometrial cancers that often present with difficult differential diagnoses. Less