Abstract

MUC5AC has been indicated to be a marker for mucinous ovarian cancer (OC). We investigated the use of in situ proximity ligation assay (PLA) for blood group ABH expressing MUC5AC to differentiate between serous and mucinous OC, to validate preceding observations that also MUC5AC ABH expression is increased in mucinous OC. We developed PLA for anti-A, B, and H/anti-MUC5AC and a PLA using a combined lectin Ulex europaeus agglutinin I (UEA I)/anti-MUC5AC assay. The PLAs were verified with mass spectrometry, where mucinous OC secretor positive patients’ cysts fluids containing ABH O-linked oligosaccharides also showed positive OC tissue PLA staining. A nonsecretor mucinous OC cyst fluid was negative for ABH and displayed negative PLA staining of the matched tissue. Using the UEA I/MUC5AC PLA, we screened a tissue micro array of 410 ovarian tissue samples from patients with various stages of mucinous or serous OC, 32 samples with metastasis to the ovaries and 34 controls. The PLA allowed differentiating mucinous tumors with a sensitivity of 84% and a specificity of 97% both against serous cancer but also compared to tissues from controls. This sensitivity is close to the expected incidence of secretor individuals in a population. The recorded sensitivity was also found to be higher compared to mucinous type cancer with metastasis to the ovaries, where only 32% were positive. We conclude that UEA 1/MUC5AC PLA allows glycospecific differentiation between serous and mucinous OC in patients with positive secretor status and will not identify secretor negative individuals with mucinous OC.

中文翻译：

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原位邻近结扎法分析粘液性卵巢癌组织中 MUC5AC 上的血型抗原

摘要

MUC5AC 已被证明是粘液性卵巢癌 (OC) 的标志物。我们研究了使用原位邻近结扎测定 (PLA) 对表达 MUC5AC 的血型 ABH 来区分浆液性和粘液性 OC，以验证先前的观察结果，即 MUC5AC ABH 表达在粘液性 OC 中也增加。我们开发了用于抗 A、B 和 H/抗 MUC5AC 的 PLA，以及使用联合凝集素欧洲葡萄凝集素 I (UEA I)/抗 MUC5AC 测定的 PLA。PLAs 用质谱法验证，其中粘液性 OC 分泌阳性患者的囊液含有 ABH O-连接的寡糖也显示阳性OC组织PLA染色。非分泌型黏液性 OC 囊液 ABH 呈阴性，匹配组织的 PLA 染色呈阴性。使用 UEA I/MUC5AC PLA，我们筛选了来自不同阶段粘液性或浆液性 OC 患者的 410 个卵巢组织样本、32 个转移至卵巢的样本和 34 个对照的组织微阵列。PLA 允许区分粘液性肿瘤，其对浆液性癌的敏感性为 84%，特异性为 97%，而且与对照组的组织相比也是如此。这种敏感性接近人群中分泌个体的预期发生率。与转移到卵巢的粘液型癌症相比，记录的敏感性也更高，其中只有 32% 是阳性的。