Metallothionein mediates the level and activity of nuclear factor κB in murine fibroblasts

被引:48
作者
Butcher, HL
Kennette, WA
Collins, O
Zalups, RK
Koropatnick, J
机构
[1] London Reg Canc Ctr, London, ON N6A 4L6, Canada
[2] Univ Western Ontario, Dept Oncol, London, ON, Canada
[3] Univ Western Ontario, Dept Microbiol & Immunol, London, ON, Canada
[4] Univ Western Ontario, Dept Pathol, London, ON, Canada
[5] Univ Western Ontario, Dept Physiol & Pharmacol, London, ON, Canada
[6] Mercer Univ, Sch Med, Div Basic Med Sci, Macon, GA 31207 USA
关键词
D O I
10.1124/jpet.104.066126
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The zinc-binding protein metallothionein (MT) is associated with resistance to apoptosis. We examined whether MT regulates the zinc-dependent antiapoptotic transcription factor nuclear factor kappaB (NF-kappaB), which is up-regulated under many conditions that lead to elevated MT expression. NF-kappaB protein levels and NF-kappaB-dependent reporter gene activity were examined in clonal MT(+) (MT-WT) and MT(-) (MT-KO) fibroblastic cell lines. The amount of cellular NF-kappaB p65 protein in MT-KO was less than 20% of the amount in MT-WT cells, in accord with increased sensitivity of MT-KO cells to apoptosis. NF-kappaB p65 mRNA levels, and NF-kappaB p50 subunit and IkappaBalpha protein levels, were unchanged. NF-kappaB activity assessed by expression of a transfected NF-kappaB reporter construct was less than half that observed in MT-KO cells. Decreased nuclear localization of NF-kappaB p65 in MT- KO clones was not responsible for differences in activity. In fact, MT- KO cells had higher nuclear levels of NF-kappaB p65 than did MT- WT cells, despite a lower cellular NF-kappaB level and function, suggesting that metallothionein mediated the specific activity of NF-kappaB. Reconstitution of MT by stable incorporation of an MT- 1 expression vector in MT- KO cells resulted in increased NF-kappaB p65 ( but not IkappaBalpha or NF-kappaB p50), increased NF-kappaB-dependent reporter activity, and increased resistance to apoptosis. These data support the hypothesis that metallothionein positively regulates the cellular level and activity of NF-kappaB.
引用
收藏
页码:589 / 598
页数:10
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