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

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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