Koh S H, Toh B H, Gallichio H A, Elrick W L
Aust J Exp Biol Med Sci. 1978 Jun;56(3):313-20. doi: 10.1038/icb.1978.33.
Tissue culture monolayers of seven human intracranial tumours comprising 2 astrocytomas, 3 meningiomas, 1 secondary squamous cell carcinoma and 1 secondary adenocarcinoma were examined by a double immunofluorescent staining technique to demonstrate Concanavalin A (Con A) surface receptors and cytoplasmic actin in the same cell. Tumour cells, treated with fluoresceinisothiocyanate-labelled Con A (FITC-Con A) showed staining in cell margins or in a random distribution over the cell surface. Incubating the cells with FITC-Con A at 37 degrees for increasing periods of time resulted first in staining of clusters and later of perinuclear globules. Cells, pretreated with 4% paraformaldehyde at 4 degrees for 10 min or with cytochalasin B at 37 degrees for 30 min showed staining restricted to cell margins. In the cytochalasin B-treated cells, the peripheral staining was in the form of coarse clusters. Double fluorochrome studies showed that the anti-actin antibody (AAA) staining occurred in sites closely related to those stained by FITC-Con A both in untreated as well as in cytochalasin B-treated cells. The findings suggest that Con A receptors, as an example of a stable cell membrane determinant in human tumour cells, are associated with actin and that their mobility on the cell surface is dependent on an intact cytoplasmic actin system.
采用双重免疫荧光染色技术，对包含2例星形细胞瘤、3例脑膜瘤、1例继发性鳞状细胞癌和1例继发性腺癌的7种人类颅内肿瘤的组织培养单层细胞进行检测，以在同一细胞中显示刀豆球蛋白A（Con A）表面受体和细胞质肌动蛋白。用异硫氰酸荧光素标记的Con A（FITC-Con A）处理的肿瘤细胞在细胞边缘或细胞表面呈随机分布染色。将细胞与FITC-Con A在37℃孵育不同时间，首先导致簇状染色，随后是核周小球染色。用4%多聚甲醛在4℃预处理10分钟或用细胞松弛素B在37℃预处理30分钟的细胞，染色仅限于细胞边缘。在细胞松弛素B处理的细胞中，周边染色呈粗大簇状。双荧光染料研究表明，抗肌动蛋白抗体（AAA）染色发生在与FITC-Con A染色部位密切相关的部位，无论是未处理的细胞还是细胞松弛素B处理的细胞。这些发现表明，Con A受体作为人类肿瘤细胞中稳定细胞膜决定因素的一个例子，与肌动蛋白相关，并且它们在细胞表面的移动性依赖于完整的细胞质肌动蛋白系统。
