Abstract:
:2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is known to affect various cellular activities including growth factor signal transduction, hormone responses, and cell differentiation. The purpose of this study was to examine more closely the very early effects of TCDD on protein tyrosine kinase activity, specifically p60(Src). We found that TCDD causes rapid changes in the plasma-microsomal membrane levels and activity of p60(Src) in Hepa 1c1c7, Hepa c4 cells, and SR3Y1 cells, a p60(v-Src) overexpressing cell line. Such cellular changes occur within 30 minutes after 10 nM TCDD treatment, as measured by Western blot analysis. TCDD's ability to increase p60(Src) levels was found to be: (1) dose-dependent, with an estimated EC(50) between 10(-10) and 10(-11) M TCDD; (2) Ah receptor (AhR)-dependent, since TCDD's effect was blocked by co-administration with 1 μM α-naphthoflavone, an AhR antagonist; and interestingly (3) ARNT-independent, since TCDD's effect was observed in Hepa c4 cells, an ARNT(-) mutant cell line. Since ARNT is a heterodimerization partner of the AhR required for binding of the ligand-activated AhR to dioxin-responsive elements on DNA in the nucleus to transactivate genes controlled by the AhR, an alternative mechanism for TCDD's action is discussed which does not require ARNT. Along with increased membrane levels of p60(Src), we observed a corresponding increase in the activity of a 60 kDa protein tyrosine kinase using two different kinase detection assays. This effect of TCDD was also found to be AhR-dependent, ARNT-independent, and independent of de novo protein synthesis since cycloheximide was unable to completely abolish TCDD's effect. The present findings provide a potentially important mechanism by which TCDD can alter cell growth and differentiation.
journal_name
Environ Toxicol Pharmacoljournal_title
Environmental toxicology and pharmacologyauthors
Blankenship A,Matsumura Fdoi
10.1016/s1382-6689(97)00016-1subject
Has Abstractpub_date
1997-07-01 00:00:00pages
211-20issue
3eissn
1382-6689issn
1872-7077pii
S1382-6689(97)00016-1journal_volume
3pub_type
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