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

开发新的诊断方法来识别子宫内膜癌细胞表达的岩藻糖基化碳水化合物链及其临床应用

• 批准号: 12557140
• 负责人: NOZAWA Shiro
• 金额: $ 8.32万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557140/
• 关键词: endometrial cancers; Cytoloaic diagnosis; Thin Prep; enzyme immunoaasay (EmC-EIA); cervical gland cells; EmC-EIA; MSN-1

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

子宫内膜癌的细胞学诊断通常存在形态学解释困难，因此可能导致假阳性诊断。本研究的目的是开发一种新的酶免疫测定法（EmC-EIA），该方法靶向子宫内膜癌特异性表达的碳水化合物抗原（SN-Ag）。从子宫内膜取样获得的细胞学标本用单层细胞制备装置“Thin Prep”处理。该方法作为子宫内膜癌辅助诊断方法的有用性在手术和临床标本上进行了评价。共获得44例子宫内膜癌患者的手术标本（G1：32例，G2：5例，G3：5例），另外还有10例正常子宫内膜标本。将细胞学标本用“Thin Prep”法粘附在滤膜上，浸入细胞解离缓冲液中，振摇后，用超声波显微分级， ...更多信息 或.使用特异性检测SN-Ag的单克隆抗体MSN-1通过夹心EIA测定样品中的SN-Ag量。可检出SN-Ag的样品确定为阳性样品。72.7%（G1：75.0%，G2：80.0%，G3：40.0%的子宫内膜癌标本EmC-EIA阳性，而12例正常子宫内膜标本的阳性率为0%。在直接观察下获得的手术标本仅包括预期的癌组织，不包括周围的正常组织或其他细胞类型。这些结果反映为与正常子宫内膜标本相比，子宫内膜标本中的高阳性率，以及正常子宫内膜标本中的0%的假阳性率，并且证明我们的EmC-EIA方法能够准确地测量细胞学标本中的SN-Ag量。在137例临床标本中，EMC-EIA检测子宫内膜标本的阳性率为69.4%，其中G1期阳性率最高，为73.11%，G2期阳性率为60.0%，G3期阳性率为60.0%，与手术标本的阳性率相近。另一方面，84例正常子宫内膜标本的阳性率为15.5%，与手术标本相比，假阳性率增加。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宫颈腺细胞所致。17例子宫腺肌病患者的阳性率为35.3%;这介于子宫内膜癌和正常子宫内膜标本的阳性率之间。因此，我们的结果表明，我们的EmC-EIA检测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，与适当的采样的子宫内膜腔，这种方法将作为一种辅助诊断方法，并将在临床上适用于评价细胞学标本的子宫内膜癌，特别是高度分化的子宫内膜癌，往往与困难的鉴别诊断。少

项目成果

久布白 兼行 et al. 4: "子宮頸部円錐切除術における妊孕脳温存"産科と婦人科. 68・8. 1024-1030 (2001)

Kaneyuki Kubushiro 等 4：“宫颈锥切术中的妊娠脑保存”妇产科 68・8（2001 年）。

Kyoko TAKEHARA et al. 5: "Expressison of an isoform of the testis-specific estrogen sulfortransferase in the rifle placenta during the late gestational period"Arch. Biochem. Biophys. 394・2. 201-208 (2001)

Kyoko TAKEHARA 等人 5：“妊娠晚期睾丸特异性雌激素硫基转移酶的表达”Biochem. 201-208。

Eiko, SAITO., et al.: "The expression of β 1, 4-garactosyltransferase in human trophoblasts"Acta Histochem.Cytochem.. 34(1). 62(A24) (2001)

Eiko，SAITO.，等人：“β1,4-半乳糖基转移酶在人滋养层中的表达”Acta Histochem.Cytochem.. 34(A24)(2001)。

Yoshibumi KANETA., et al.: "Selective cytotoxicity of adriamycin immunoconjugate of monoclonal antibody MSN-1 to endometrial adenocarcinoma in vitro and in vivo"Oncol.Rep.. 7. 1099-1106 (2000)

Yoshibumi KANETA.等：“单克隆抗体MSN-1的阿霉素免疫缀合物对子宫内膜腺癌的体外和体内选择性细胞毒性”Oncol.Rep..7.1099-1106(2000)

Kaneyuki Kubushiro, et al 6: "Biological characteristics of human uterine endometrial cancer variant cells selected for blood group H type 1 antigen : adhesion to vascular endothelial cells."Acta Histocemica et Cytochemica. 33・3. 209-213 (2000)

Kaneyuki Kubushiro等6：“针对H型1型抗原选择的人子宫内膜癌变异细胞的生物学特性：与血管内皮细胞的粘附”。 Acta Histochemica et Cytochemica 33·3（2000）。