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cAMP‐induced phenotypic reversion of adhesion, aggregation, and endocytosis in adhesion‐defective CHO cell variants
Author(s) -
Cheung E.,
Juliano R. L.
Publication year - 1985
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041240225
Subject(s) - fibronectin , incubation , endocytosis , adhesion , population , cell adhesion , cell , phosphodiesterase , microbiology and biotechnology , cell culture , fetal bovine serum , biology , bovine serum albumin , chinese hamster ovary cell , in vitro , chemistry , receptor , biochemistry , enzyme , medicine , genetics , environmental health , organic chemistry
AD v F11 cells are a CHO adhesion variant which, unlike wild type (WT) cells, are not able to adhere to fibronectin (Fn) coated substrata or to be aggregated by Fn‐beads. However, AD v F11 cells bind Fn‐beads to the same extent as WT cells, thus suggesting that the defect(s) associated with AD v F11 cells are distal to the initial receptor‐ligand binding event (Cheung and Juliano, Exp. Cell Res. 152:127, 1984). In this communication we report that cAMP analogs such as dibutyryl‐cAMP (dbcAMP) and 8‐bromo‐cAMP are able to correct defect(s) associated with AD v F11 cells enabling them to adhere to Fn‐coated dishes and to aggregate in the presence of Fn‐beads. However, only approximately 40% of AD v F11 cells were found to be responsive to dbcAMP suggesting heterogeneity in the cell population with respect to dbcAMP sensitivity. Further analysis of this partial response led us to isolate a subclone of AD v F11 cells, F11CA11, which is highly responsive to dbcAMP treatment. Induction of Fn‐mediated cell adhesion and aggregation in F11CA11 by dbcAMP is both time and dose dependent. Optimal responses were obtained after overnight incubation in alpha‐MEM containing, 1% fetal calf serum, 4% bovine serum albumin, 0.5 mM dbcAMP and 0.2 mM methyl‐isobutyl‐xanthine (MIX), a phosphodiesterase inhibitor. Under these conditions, 70–80% of F11CA11 cells were found to be adherent, compared to 5–7% of untreated F11CA11 cells and 95–100% of WT cells. Aggregation of dbcAMP‐MIX treated F11CA11 cells induced by Fn‐beads also approached that of WT cells. In addition, treatment with dbcAMP‐MIX markedly increased the ability of F11CA11 cells to internalize Fn‐beads. The maintenance of the adherent phenotype required the constant presence of dbcAMP‐MIX. Removal of dbcAMP‐MIX from the incubation medium resulted in return to the original nonadhesive phenotype. Thus, elevation of cAMP levels can dramatically modify the behavior of F11CA11 cells with respect to fibronectin mediated adhesion, aggregation and endocytosis, in effect causing a phenotypic reversion of all three parameters to wild type status. This suggests that the mechanisms for adhesion, aggregation and endocytosis may each involve regulation by cyclic AMP‐protein kinase systems.