TY - JOUR
T1 - Antisense BAG-1 sensitizes HeLa cells to apoptosis by multiple pathways
AU - Xiong, Jieying
AU - Chen, Jun
AU - Chernenko, Garry
AU - Beck, Jessalyn
AU - Liu, Hongyu
AU - Pater, Alan
AU - Tang, Shou-Ching
N1 - Funding Information:
We thank the Canadian Institutes of Health Research for sponsoring our work through the Canadian Breast Cancer Research Initiative Grant 338581.
PY - 2003/12/19
Y1 - 2003/12/19
N2 - To study the mechanism of action of BAG-1 in drug-induced apoptosis, we constructed an antisense BAG-1 vector and established a stably transfected cell line from BAG-1-over-expressing HeLa cells. Reduced BAG-1 protein was confirmed by Western blot. Treatment of the antisense BAG-1-transfected cells with the anti-cancer drugs staurosporine, paclitaxel, all-trans retinoic acid (ATRA), and N-(4-hydroxyphenyl) retinamide (4-HPR) resulted in significantly enhanced apoptosis and reduced cell viability relative to vector-transfected cells. While the expression of p53 was increased, the level of Bcl-2 and Bax was decreased. Cells underexpressing BAG-1 had reduced cytosolic cytochrome c level. Treatment with staurosporine and paclitaxel resulted in increased cytochrome c release from mitochondria, whereas there was no change induced by treatment with ATRA and 4-HPR. Our experiments suggest that BAG-1 inhibits anti-cancer drug-induced apoptosis through apoptosis regulation pathways that may involve the mitochondrial Bcl-2/Bax ratio, p53, and differential anti-cancer drug-mediated cytochrome c release.
AB - To study the mechanism of action of BAG-1 in drug-induced apoptosis, we constructed an antisense BAG-1 vector and established a stably transfected cell line from BAG-1-over-expressing HeLa cells. Reduced BAG-1 protein was confirmed by Western blot. Treatment of the antisense BAG-1-transfected cells with the anti-cancer drugs staurosporine, paclitaxel, all-trans retinoic acid (ATRA), and N-(4-hydroxyphenyl) retinamide (4-HPR) resulted in significantly enhanced apoptosis and reduced cell viability relative to vector-transfected cells. While the expression of p53 was increased, the level of Bcl-2 and Bax was decreased. Cells underexpressing BAG-1 had reduced cytosolic cytochrome c level. Treatment with staurosporine and paclitaxel resulted in increased cytochrome c release from mitochondria, whereas there was no change induced by treatment with ATRA and 4-HPR. Our experiments suggest that BAG-1 inhibits anti-cancer drug-induced apoptosis through apoptosis regulation pathways that may involve the mitochondrial Bcl-2/Bax ratio, p53, and differential anti-cancer drug-mediated cytochrome c release.
KW - Antisense down-regulation
KW - Apoptosis
KW - BAG-1
KW - Bcl-2 apoptotic family
KW - Mitochondrial pathway
KW - p53
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U2 - 10.1016/j.bbrc.2003.10.160
DO - 10.1016/j.bbrc.2003.10.160
M3 - Article
C2 - 14680805
AN - SCOPUS:0344121269
SN - 0006-291X
VL - 312
SP - 585
EP - 591
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -