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人结肠癌细胞(Caco-2)单层对表皮生长因子的结合

Binding of epidermal growth factor by human colon carcinoma cell (Caco-2) monolayers.

作者信息

Hidalgo I J, Kato A, Borchardt R T

机构信息

Department of Pharmaceutical Chemistry, School of Pharmacy, University of Kansas, Lawrence 66045.

出版信息

Biochem Biophys Res Commun. 1989 Apr 14;160(1):317-24. doi: 10.1016/0006-291x(89)91658-6.

Abstract

The objective of this study was to investigate whether Caco-2 cells bind and internalize epidermal growth factor (EGF). [125I]EGF was presented to the apical (AP) or basolateral (BL) side of Caco-2 monolayers, grown on microporous membranes, at different times in culture. At day 10, [125I]EGF binding (at 37 degrees C) to the BL membrane was 2-3 times greater than binding to the AP membrane. Of that [125I]EGF bound to the AP membrane 76% was internalized within 3 h while internalization from the BL membrane was 90%. At lower temperatures membrane-bound [125I]EGF increased while internalization decreased. At day 16, AP and BL binding decreased and then remained constant through day 25. [125I]EGF was bound to the BL membrane of 10 days old monolayers with a Kd of 0.67 nM. There was a single binding site whose numbers in the BL membrane was about 5500/cell.

摘要

本研究的目的是调查Caco-2细胞是否结合并内化表皮生长因子(EGF)。将[125I]EGF在培养的不同时间呈现给生长在微孔膜上的Caco-2单层细胞的顶端(AP)或基底外侧(BL)面。在第10天,[125I]EGF(在37℃)与BL膜的结合比与AP膜的结合大2至3倍。结合到AP膜上的[125I]EGF中,76%在3小时内被内化,而从BL膜内化的比例为90%。在较低温度下,膜结合的[125I]EGF增加,而内化减少。在第16天,AP和BL结合减少,然后在第25天保持恒定。[125I]EGF与10天大的单层细胞的BL膜结合,解离常数(Kd)为0.67 nM。存在一个单一的结合位点,其在BL膜中的数量约为5500/细胞。

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