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• W2967651168 abstract "5202 Certain forms of hexavalent chromium [Cr (VI)] are human carcinogens. Previous work in our lab has shown that a broad range protein tyrosine phosphatase (PTP) inhibitor, sodium orthovanadate (SOV) partially abrogated Cr (VI) - induced growth arrest and induced cell cycle checkpoint override, which was coincident with Akt activation. The aim of the present study was to determine the correlation between the Cr (VI)-induced Akt downregulation on the localization and expression of various effectors of the G1/S checkpoint (p21, p27, Cyclin D1 and phospho Rb) and the role of PTP inhibition in this effect. HLFs were treated with 0 -6µM of Cr (VI), with and without 10µM SOV, fixed on 8-well chambered coverslips and incubated with a primary antibody to either p21, p27, Cyclin D1 or phospho Ser 807/811 Rb followed by treatment with a secondary antibody conjugated to a fluorochrome, Alexafluor 488. These experiments were also performed with HLFs transfected with Akt-KD (kinase dead) and constitutively active Myr-Akt plasmids as well as HLFs in which Akt1 was silenced with siRNA. Cr (VI) caused nuclear localization of p27 as early as 4h after exposure. The addition of SOV alone had no effect on nuclear localization, but caused the nuclear export of p27 in the presence of Cr (VI), which was maximum at 24 hours. In cells in which Akt was knocked down by siRNA or transfected with the Akt-KD plasmid, p27 was localized in the nucleus after Cr(VI) exposure, even in the presence of SOV. Conversely, in cells transfected with the Myr-Akt plasmid, there was no nuclear localization of p27 in the presence of Cr(VI). Finally, Cr(VI) induced a punctuate nuclear staining of pRb which was abrogated by PTP inhibition. The total cellular levels of p27, p21, Cyclin D1, pRb and pSer473 Akt were determined by Western blotting techniques. Cr (VI) induced a dose-dependant decrease in the expression of p27, pSer 473 Akt, Cyclin D1 and pRb after 24h exposure, while SOV abrogated a decrease only in pSer 473 Akt and pRb expression. In conclusion, Akt activation is necessary for the cytosolic retention of p27 by SOV, which may be responsible in part for the previously observed activity of this PTP inhibitor in overriding Cr (VI)-induced checkpoint arrest and this effect appears to be independent of p27 protein expression. The bypass of cell cycle checkpoint arrest by Akt activation in the face of genotoxic damage could increase genomic instability and metastasis, which is a hallmark of neoplastic progression. This study was supported by NIH grants CA107972 to SC and E505304 and E509961 to SRP." @default.
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• W2967651168 date "2007-05-01" @default.
• W2967651168 modified "2023-09-27" @default.
• W2967651168 title "Protein tyrosine phosphatase inhibition abrogates the effect of Cr(VI) on the effectors of the G1/S checkpoint through mechanisms involving Akt" @default.
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