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Promotion-resistant JB6 mouse epidermal cells exhibit defects in phosphatidylethanolamine synthesis and phorbol ester-induced phosphatidylcholine hydrolysis.

ABSTRACT: The tumour-promotion-sensitive (P+) and -resistant (P-) variants of mouse JB6 epidermis-derived cells have often been used to study the requirements for the tumour-promoting effect of PMA. As part of an effort to identify the defect(s) in JB6 P- cells that might prevent the promoting effect of PMA, stimulation of phospholipase D (PLD)-mediated hydrolysis of phosphatidylcholine (PtdCho) and phosphatidylethanolamine (PtdEtn) by PMA as well as the rate of phospholipid synthesis were compared in three P+ variants, two P- variants and a transformed variant of the JB6 cell line. PMA (5-100 nM) had significantly less stimulatory effect on PtdCho hydrolysis in P- cells than in P+ or transformed JB6 cells. The effects of PMA on PtdEtn hydrolysis in the P+ and P- cell lines were similar, whereas in transformed cells PMA had slightly less effect. Each JB6 cell line was found to contain similar amounts of PtdCho. In contrast, P- cells contained significantly less PtdEtn and a correspondingly higher level of ethanolamine phosphate compared with P+ and transformed cells. P- cells also secreted ethanolamine phosphate into the medium; this process was greatly enhanced by PMA. In the two P- variants the synthesis of PtdEtn from [14C]ethanolamine was reduced to various extents, whereas the rate of PtdCho synthesis was comparable in each JB6 cell line. The synthesis of PtdCho, but not PtdEtn, was greatly stimulated by PMA in both the P+ and P- clones. The results indicate that decreased synthesis/level of PtdEtn and suboptimal functioning of a PtdCho-specific PLD are common characteristics of the P- JB6 cells examined so far. The observed alterations in phospholipid metabolism may play a role in the resistance of P- cells to the tumour-promoting action of PMA.

PROVIDER: S-EPMC1218346 | BioStudies | 1997-01-01

REPOSITORIES: biostudies

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