Abstract:
:In vivo and in vitro studies performed on the polar solvent N-methylformamide (NMF), as well as on its association with chemotherapeutic agents or X rays, have clearly demonstrated that this compound is capable of inducing changes in biological characteristics of tumor cells, e.g., cell differentiation. However, the mechanism of action of NMF is far from being elucidated. Hence, in order to better clarify such a mechanism an in vitro study was carried out by using mouse fibroblasts in primary culture (MEF) and human melanoma cultured cells (M14). Results obtained by immunocytochemical and ultrastructural methods with doses of NMF ranging from 0.1 to 7% are reported here. As a general rule, a different sensitivity (in terms of cytopathologic changes induced by NMF) was found between the cell types considered. In fact, melanoma cells appeared to be highly susceptible to the action of the drug, undergoing severe morphological modifications represented mainly by a reversible dose and time-dependent cell rounding and surface blebbing. In contrast, NMF-induced injury in MEF cells was characterized mainly by a simple retraction of the cell body. A cytochemical analysis of the expression of certain membrane antigens (e.g., glycoproteins, epidermal growth factor receptor, B2 microglobulin) in NMF-treated M14 cells undergoing blebbing was also carried out. A randomly distributed labeling of such molecules was observed. Accordingly, freeze-fracturing electron microscopic analysis also displayed a random distribution of intramembrane particles over the plasma membrane. When subcellular changes induced by the drug were investigated, a remarkable modification of cytoskeletal components was detected in both cell types. In particular, cross-linked actin microfilament bundles were easily observed in NMF-exposed MEF cells. Finally, when different experimental conditions which perturb calcium ion homeostasis or restore protein thiol group reduced state were analyzed, a noticeable impairment of the blebbing phenomenon was observed. Thus, a target effect of NMF on the microfilament system, probably leading, in turn, to several subcellular changes and cell surface blebbing, can be hypothesized. Such a cytoskeletal element-dependent cytopathology appears to be related to changes of the oxidized state of such molecules as well as to calcium ion perturbations.
journal_name
Exp Mol Patholjournal_title
Experimental and molecular pathologyauthors
Malorni W,Meschini S,Arancia Gdoi
10.1016/0014-4800(92)90001-rkeywords:
subject
Has Abstractpub_date
1992-10-01 00:00:00pages
85-104issue
2eissn
0014-4800issn
1096-0945pii
0014-4800(92)90001-Rjournal_volume
57pub_type
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