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Reconstitution of galectin-3 alters glutathione content and potentiates TRAIL-induced cytotoxicity by dephosphorylation of Akt
Journal article   Peer reviewed

Reconstitution of galectin-3 alters glutathione content and potentiates TRAIL-induced cytotoxicity by dephosphorylation of Akt

Yong J Lee, Young K Song, Jae J Song, R.Rita Siervo-Sassi, Hyeong-Reh C Kim, Ling Li, Douglas R Spitz, Anna Lokshin and Jin H Kim
Experimental cell research, Vol.288(1), pp.21-34
2003
DOI: 10.1016/S0014-4827(03)00211-8
PMID: 12878156

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Abstract

We investigated the role of galectin-3 in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptotic death in human breast carcinoma BT549 cells. We observed that parental galectin-3 null BT549 cells (BT549par) as well as control vector transfected (BT549neo) cells were resistant to TRAIL, while galectin-3 cDNA-transfected BT549 cells (BT549gal-3) were sensitive to TRAIL. Data from flow cytometry and immunoblotting analyses reveal that reconstitution of galectin-3 promoted cell death and PARP cleavage as well as caspase (-8, -9, and -3) activation during TRAIL treatment. However, unlike TRAIL treatment, galectin-3 transfectants were resistant to UV-B-induced PARP cleavage. Data from cDNA array analysis show that galectin-3 did not significantly enhance or reduce any apoptosis-related gene expression. Moreover, although galectin-3 restored pre-mRNA splicing activity and resulted in elevation of FLIPs protein, experiments with FLIPs cDNA-transfected cells show that overexpression of FLIPs did not sensitize cells to TRAIL. Interestingly, BT549gal-3 cells demonstrated a ∼2-fold increase in total glutathione content as well as a ∼5-fold increase in GSSG content in comparison to BT549par and BT549neo cells, suggesting that galectin-3 overexpression may alter intraceullular oxidation/reduction reactions affecting the metabolism of glutathione and other thiols. In addition, galectin-3 overexpression inactivated Akt by dephosphorylation. Finally, overexpression of constitutively activated Akt protected BT549gal-3 cells from TRAIL-induced cytotoxicity. Taken together, our data suggest that galectin-3-enhanced TRAIL-induced cytotoxicity is mediated through dephosphorylation of Akt, possibly through a redox-dependent process.
Apoptosis Akt cDNA array FLIPs Galectin-3 TRAIL

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