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培养的卵巢支持细胞瘤和卵泡颗粒细胞的凝集素结合位点。

Lectin binding sites of cultured ovarian Sertoli cell tumors and follicular granulosa cells.

作者信息

Stoica G, Sowa B A

机构信息

Department of Veterinary Pathology, Texas A&M University, College Station 77840.

出版信息

Anticancer Res. 1989 May-Jun;9(3):687-94.

PMID:2764515
Abstract

A panel of seven alkaline phosphatase labeled lectins was used to probe nitrocellulose electroblots of SDS-PAGE separated proteins from a primary culture of normal ovarian granulosa cells and an ENU-induced Sertoli cell tumor cell line (SCTL-I). Several additional lectin binding proteins were observed in silver stained SDS-PAGE gels as well as with lectins in SCTL-I. Succinated concanavalin A (Suc. Con A), Ricin communis agglutinin (RCA-I), Ulex europaeus agglutinin (UEA 1), Soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA) and Peanut agglutinin (PNA) stained more intensely in SCTL-I than normal granulosa cells. The same lectins as above, labeled with fluorescein isothiocyanate (FITC), were used to study the distribution of specific binding sites of tissue cultured cells grown in chamber/slides. Both normal ovarian granulosa cells and SCT cells exhibited strong peninuclear cytoplasmic labeling with Con A UEA-1 and WGA exhibited predominantly a nuclear and granular cytoplasmic staining pattern. SBA and DBA exhibited a strong coarse granular cytoplasmic labeling in granulosa cells and moderate granular cytoplasmic in SCT cells. In granulosa cells, Golgi regions stained strongly with PNA but weakly in SCT cells. RCA-I staining was negative in both cultures. Labeling of tissue cultured cells with lectins provides more details than histological sections of lectins binding sites at cellular structural levels.

摘要

使用一组七种碱性磷酸酶标记的凝集素来探测经SDS-PAGE分离的正常卵巢颗粒细胞原代培养物和ENU诱导的支持细胞瘤细胞系(SCTL-I)中的蛋白质的硝酸纤维素膜电印迹。在银染的SDS-PAGE凝胶以及SCTL-I中的凝集素中还观察到几种额外的凝集素结合蛋白。琥珀酰化伴刀豆球蛋白A(Suc. Con A)、蓖麻凝集素(RCA-I)、欧洲荆豆凝集素(UEA 1)、大豆凝集素(SBA)、双花扁豆凝集素(DBA)和花生凝集素(PNA)在SCTL-I中的染色比正常颗粒细胞更强烈。使用上述相同的用异硫氰酸荧光素(FITC)标记的凝集素来研究在培养室/载玻片上生长的组织培养细胞的特异性结合位点的分布。正常卵巢颗粒细胞和SCT细胞均表现出伴刀豆球蛋白A、UEA-1的强烈核周细胞质标记,而WGA主要表现出核和颗粒状细胞质染色模式。SBA和DBA在颗粒细胞中表现出强烈的粗颗粒状细胞质标记,在SCT细胞中表现出中等颗粒状细胞质标记。在颗粒细胞中,高尔基体区域用PNA染色强烈,但在SCT细胞中染色较弱。两种培养物中RCA-I染色均为阴性。用凝集素标记组织培养细胞比在细胞结构水平上对凝集素结合位点进行组织学切片提供了更多细节。

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