Increased genotoxicity of acetylaminofluorene by modulators of multixenobiotic resistance mechanism: studies with the fresh water clam Corbicula fluminea.
The presence of a 'multixenobiotic resistance' [MXR] mechanism in gills of the freshwater clam Corbicula fluminea was investigated. Western blot analyses of membrane vesicles from gills, applying antibodies to vertebrate P170 multidrug resistance (MDR) protein, revealed a 135 kDa immunoreactive protein. Verapamil caused a reduction of 3H-vincristine (3H-VCR) binding onto vesicles from clam. Exposure of clams to 3H-VCR in the presence of verapamil or staurosporine (STP) enhanced the accumulation of 3H-VCR over control values. Furthermore, clams were exposed instead to VCR, to a model carcinogen, 2-acetylaminofluorene (AAF), to determine the verapamil- and STP-dependent increase of single-strand breaks (SSBs) in DNA from gills of this organism. Verapamil caused no or little increase of SSBs induced by exposure to 0.01 or 0.10 microM AAF, respectively, as measured by the alkaline elution technique. In contrast, in the presence of STP a highly significant and dose-dependent enhancement of AAF-mediated SSBs was measured already at exposure to 0.01 microM AAF. These data indicate (i) that the clam C. fluminea is provided with a P-glycoprotein-like element of the MDR-mechanism, (ii) that this system can be poisoned by chemosensitizers such as verapamil and STP, (iii) the role of protein kinase C in the regulation of MXR function and (iv) the importance of the MXR modulators for the assessment of ecotoxicological effects of pollutants.
Waldmann
P
P
Ruder Boskovic Institute, Center for Marine Research, Zagreb, Croatia.
Pivcevic
B
B
Müller
W E
WE
Zahn
R K
RK
Kurelec
B
B
D015073:2-Acetylaminofluorene / Q000633:toxicity*; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis*; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D049872:Bivalvia / Q000187:drug effects* / Q000235:genetics; D015153:Blotting, Western; D004249:DNA Damage; D018432:Drug Resistance, Multiple; D005880:Gills / Q000187:drug effects; D009153:Mutagens / Q000633:toxicity*; D019311:Staurosporine; D014700:Verapamil / Q000494:pharmacology; D014750:Vincristine / Q000378:metabolism; D015262:Xenobiotics / Q000494:pharmacology*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000470:Alkaloids; D009153:Mutagens; D015262:Xenobiotics; D014750:Vincristine; D015073:2-Acetylaminofluorene; D014700:Verapamil; D019311:Staurosporine