Poly (ADP-ribose) polymerase activity regulates apoptosis in HeLa cells after alkylating DNA damage.
Majority of chemotherapeutic agents inhibit tumor growth by inducing apoptosis or necrosis. The DNA alkylating agent, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), kills cells by necrosis through massive production of DNA strand breaks and subsequent over-activation of PARP. Inhibition of PARP, either through PARP1 genetic ablation or through small molecule PARP inhibitors, protected MNNG-induced cell death in certain cell types including MEF and primary cortical cultures. We report here that a potent PARP inhibitor, ABT-888, facilitates the induction of apoptotic cell death in HeLa cells treated with MNNG. Although the release of cytochrome c from mitochondria to cytosol was observed in HeLa cells treated with either MNNG alone or the combination of MNNG and ABT-888 (MNNG/ABT-888), apoptosis is observed only in HeLa cells treated with MNNG/ABT-888. Bcl-2 family proteins regulate the release of cytochrome c. Downregulation of Bax and Bak by their corresponding siRNAs or overexpression of Bcl-xl inhibited the release of cytochrome c from mitochondria to cytosol, and inhibited apoptosis induced by MNNG/ABT-888. Further examination indicates that ATP concentration is greatly reduced in HeLa cells treated with MNNG alone, but not in HeLa cells treated with MNNG/ABT-888. Reduction of ATP concentration by F0F1-ATP synthase inhibitor oligomycin A renders HeLa cells resistant to the apoptosis induction by treatment with MNNG/ABT-888. Unlike in HeLa cells, ABT-888 protected MNNG induced cell death in normal human fibroblasts. Our study provides evidence that PARP activity determines the fate of HeLa cells by regulating the level of ATP after treatment with MNNG.
Liu
Xuesong
X
Department R47S, Cancer Research, Abbott Laboratories, Abbott Park, Illinois 60064, USA. xuesong.liu@abbott.com
Luo
Xu
X
Shi
Yan
Y
Zhu
Gui-Dong
GD
Penning
Thomas
T
Giranda
Vincent L
VL
Luo
Yan
Y
D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D001562:Benzimidazoles / Q000494:pharmacology; D045304:Cytochromes c / Q000378:metabolism; D004249:DNA Damage; D005347:Fibroblasts / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006367:HeLa Cells; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D051379:Mice; D009243:NAD / Q000737:chemistry; D009840:Oligomycins / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*
D000477:Alkylating Agents; D001562:Benzimidazoles; D009840:Oligomycins; C521013:veliparib; C031004:oligomycin A; D009243:NAD; D008769:Methylnitronitrosoguanidine; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases