Mori O, Miyasato M, Karashima T, Hashimoto T
Department of Dermatology, Kurume University School of Medicine, Japan.
J Dermatol. 1998 Aug;25(8):497-502. doi: 10.1111/j.1346-8138.1998.tb02443.x.
Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramamary Paget's disease investigated whether Paget cells proliferate as other malignant cells do. Although epithelial monolayers from the involved skin were maintained for approximately 45 days, no permanent cell lines were established. The proportion of carcinoembryonic antigen (CEA)-positive cells did not increase in the long-term cultures. Herein, we report studies of whether there is a real reduction of Paget cell numbers or if this is merely a decrease in the expression of CEA by the cells. Furthermore, we investigated whether Paget cells survive longer when cultured free from any potential inhibitory keratinocytes or other epidermal cells. Skin samples were obtained from one patient with mammary Paget's disease and three with extramammary Paget's disease; epidermal cells were cultured in vitro. An enrichment of Paget cells was carried out from the cultured epidermal cells by combining an antiepithelial membrane antigen monoclonal antibody, binding to immunobeads, and density gradient centrifugation in Nycodenz. The separated cells were re-cultured in Keratinocyte-SFM serum-free media. The proportion of CEA-positive cells did not increase in the culture, and the purified cells did not show any increase in survival times compared to the non-purified cultured cells. These results suggest that the decrease of CEA-positive cells noted during culture results from a decline in expression of CEA in the Paget cells. Paget cells in the involved epidermis do not proliferate significantly and thus differ from many other malignant cells.
先前使用来自乳腺和乳腺外佩吉特病患者受累表皮的上皮细胞原代单层培养物进行的研究，调查了佩吉特细胞是否像其他恶性细胞一样增殖。尽管来自受累皮肤的上皮单层培养物维持了约45天，但未建立永久性细胞系。癌胚抗原（CEA）阳性细胞的比例在长期培养中并未增加。在此，我们报告关于是否存在佩吉特细胞数量真正减少，或者这仅仅是细胞中CEA表达降低的研究。此外，我们研究了佩吉特细胞在无任何潜在抑制性角质形成细胞或其他表皮细胞的情况下培养时是否存活更长时间。从一名乳腺佩吉特病患者和三名乳腺外佩吉特病患者获取皮肤样本；将表皮细胞进行体外培养。通过结合与免疫磁珠结合的抗上皮膜抗原单克隆抗体以及在Nycodenz中进行密度梯度离心，从培养的表皮细胞中富集佩吉特细胞。将分离的细胞在无血清角质形成细胞培养基（Keratinocyte-SFM）中重新培养。培养物中CEA阳性细胞的比例没有增加，并且与未纯化的培养细胞相比，纯化的细胞在存活时间上没有任何增加。这些结果表明，培养过程中观察到的CEA阳性细胞减少是由于佩吉特细胞中CEA表达下降所致。受累表皮中的佩吉特细胞不会显著增殖，因此与许多其他恶性细胞不同。
