Mori O, Karashima T, Matsuo K, Hashimoto T
Department of Dermatology, Kurume University School of Medicine, Japan.
Kurume Med J. 1997;44(3):165-9. doi: 10.2739/kurumemedj.44.165.
In involved epidermis, Paget cells are completely enclosed by the surrounding keratinocytes, which appear to be intact and unaltered. It is possible that the surrounding keratinocytes inhibit Paget cell proliferation. Accordingly, Paget cells might proliferate differently when cultured as an epidermal cell suspension. In this study, primary monolayer cultures of epithelial cells from involved epidermis of patients with mammary and extramammary Paget's disease were carried out to investigate whether Paget cells proliferate in the same manner as other malignant cells. Skin samples were obtained from one patient with mammary Paget's disease, and from 2 patients with extramammary Paget's disease. The epidermis was separated from the dermis with dispase, and epidermal cell suspensions were obtained with ethylenediamine tetraacetate and trypsin. A commercially available serum-free media, Keratinocyte-SFM, was used. Epithelial monolayers from the involved skin could be maintained for approximately 45 days, while keratinocytes from normal skin were maintained for approximately 35 days. The mechanism for the longer survival of the mixed cell culture of keratinocytes and Paget cells is not known. Permanent cell lines were not developed from these primary cultures. Paget cells could not be distinguished from keratinocytes by phase-contrast microscopy. The proportion of carcinoembryonic antigen (CEA) positive cells in the culture did not increase, but instead decreased. In certain areas of the dish, the CEA positive cells proliferated and accumulated like mushrooms. However, at the periphery of the dish, the Paget cells identified by immunostaining for CEA were dispersed and not clustered. These findings indicate that the influence of keratinocytes on Paget cells also occurs in cultured cells, which may explain why Paget cells survive longer than keratinocytes. In conclusion, the Paget cells in the involved epidermis do not proliferate like other malignant cells.
在受累表皮中，派杰氏细胞完全被周围的角质形成细胞所包围，这些角质形成细胞看起来完整且未改变。周围的角质形成细胞有可能抑制派杰氏细胞的增殖。因此，当作为表皮细胞悬液培养时，派杰氏细胞的增殖方式可能会有所不同。在本研究中，对乳腺和乳腺外派杰氏病患者受累表皮的上皮细胞进行了原代单层培养，以研究派杰氏细胞是否与其他恶性细胞以相同方式增殖。从1例乳腺派杰氏病患者和2例乳腺外派杰氏病患者获取皮肤样本。用Dispase将表皮与真皮分离，并用乙二胺四乙酸和胰蛋白酶获得表皮细胞悬液。使用市售的无血清培养基角质形成细胞无血清培养基（Keratinocyte-SFM）。受累皮肤的上皮单层可维持约45天，而正常皮肤的角质形成细胞可维持约35天。角质形成细胞和派杰氏细胞混合细胞培养存活时间更长的机制尚不清楚。这些原代培养未建立起永久细胞系。通过相差显微镜无法将派杰氏细胞与角质形成细胞区分开来。培养物中癌胚抗原（CEA）阳性细胞的比例没有增加，反而下降。在培养皿的某些区域，CEA阳性细胞像蘑菇一样增殖并聚集。然而，在培养皿周边，通过CEA免疫染色鉴定的派杰氏细胞是分散的，没有聚集。这些发现表明角质形成细胞对派杰氏细胞也有影响，这可能解释了为什么派杰氏细胞比角质形成细胞存活时间更长。总之，受累表皮中的派杰氏细胞不像其他恶性细胞那样增殖。
