Latella G, Fonti R, Caprilli R, Marcheggiano A, Magliocca F, Das K M, Gambús G, Sambuy Y
Cattedra di Gastroenterologia, Università de L'Aquila, Italy.
Int J Colorectal Dis. 1996;11(2):76-83. doi: 10.1007/BF00342464.
Mature goblet cells filled with mucin ready for secretion represent about one third of the cells in primary cultures of human colonocytes. In the present study characterization of the mucins produced by cultured human colonocytes was made by histochemical methods by lectin and monoclonal antibody binding. Two monoclonal antibodies and three lectins (Dolichos biflorus (DBA), Helix pomatia (HPA) and Arachis hypogea (PNA) recognizing epitopes or sugar haptens characteristic of different stages of mucin glycoprotein maturation, were employed. The reactivity to these probes was tested both on cultured colonocytes and on tissue sections of the normal colon mucosa. The results show that the mucins produced in culture are glycosylated to the mature form, as they show the same reactivity to lectins and antibodies of the mucins expressed in tissue sections of the normal colon mucosa. In addition, it is demonstrated that cultured human colonocytes do not express mucins reactive to PNA, which are characteristic of tumors. Since the cultured colonocytes maintain the expression of differentiated functions for at least three days, they may offer a useful model to study metabolism, function and regulation of colon mucins in health and disease.
充满黏液素、随时准备分泌的成熟杯状细胞约占人结肠上皮细胞原代培养物中细胞总数的三分之一。在本研究中,通过凝集素和单克隆抗体结合的组织化学方法对培养的人结肠上皮细胞产生的黏液素进行了表征。使用了两种单克隆抗体和三种凝集素(双花扁豆凝集素(DBA)、圆口螺凝集素(HPA)和花生凝集素(PNA)),它们识别黏液素糖蛋白成熟不同阶段的表位或糖半抗原。在培养的结肠上皮细胞和正常结肠黏膜组织切片上测试了对这些探针的反应性。结果表明,培养物中产生的黏液素被糖基化至成熟形式,因为它们对正常结肠黏膜组织切片中表达的黏液素的凝集素和抗体显示出相同的反应性。此外,还证明培养的人结肠上皮细胞不表达对PNA有反应的黏液素,而这种黏液素是肿瘤的特征。由于培养的结肠上皮细胞至少三天维持分化功能的表达,它们可能为研究健康和疾病状态下结肠黏液素的代谢、功能和调节提供一个有用的模型。