An NQO1- and PARP-1-mediated cell death pathway induced in non-small-cell lung cancer cells by β-lapachone

Erik A. Bey, Melissa S. Bentle, Kathryn E. Reinicke, Ying Dong, Chin Rang Yang, Luc Girard, John D. Minna, William G. Bornmann, Jinming Gao, David A. Boothman

Research output: Contribution to journalArticle

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Abstract

Lung cancer is the number one cause of cancer-related deaths in the world. Patients treated with current chemotherapies for non-small-cell lung cancers (NSCLCs) have a survival rate of ≈15% after 5 years. Novel approaches are needed to treat this disease. We show elevated NAD(P)H:quinone oxidoreductase-1 (NQO1) levels in tumors from NSCLC patients. β-Lapachone, an effective chemotherapeutic and radiosensitizing agent, selectively killed NSCLC cells that expressed high levels of NQO1. Isogenic H596 NSCLC cells that lacked or expressed NQO1 along with A549 NSCLC cells treated with or without dicoumarol, were used to elucidate the mechanism of action and optimal therapeutic window of β-lapachone. NSCLC cells were killed in an NQO1-dependent manner by β-lapachone (LD50, ≈4 μM) with a minimum 2-h exposure. Kinetically, β-lapachone-induced cell death was characterized by the following: (i) dramatic reactive oxygen species (ROS) formation, eliciting extensive DNA damage; (ii) hyperactivation of poly(ADP-ribose)polymerase-1 (PARP-1); (iii) depletion of NAD+/ATP levels; and (iv) proteolytic cleavage of p53/PARP-1, indicating μ-calpain activation and apoptosis. β-Lapachone-induced PARP-1 hyperactivation, nucleotide depletion, and apoptosis were blocked by 3-aminobenzamide, a PARP-1 inhibitor, and 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM), a Ca2+ chelator. NQO1- cells (H596, IMR-90) or dicoumarol-exposed NQO1+ A549 cells were resistant (LD50, >40 μM) to ROS formation and all cytotoxic effects of β-lapachone. Our data indicate that the most efficacious strategy using β-lapachone in chemotherapy was to deliver the drug in short pulses, greatly reducing cytotoxicity to NQO1- "normal" cells. β-Lapachone killed cells in a tumor-selective manner and is indicated for use against NQO1+ NSCLC cancers.

Original languageEnglish (US)
Pages (from-to)11832-11837
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number28
DOIs
StatePublished - Jul 10 2007

Fingerprint

Non-Small Cell Lung Carcinoma
Cell Death
Dicumarol
Lethal Dose 50
NAD
Reactive Oxygen Species
Neoplasms
Apoptosis
Radiation-Sensitizing Agents
Drug Therapy
Ethane
Calpain
Chelating Agents
Poly (ADP-Ribose) Polymerase-1
DNA Damage
Lung Neoplasms
Oxidoreductases
Esters
Survival Rate
Nucleotides

Keywords

  • μ-calpain cell death
  • Apoptosis
  • DNA repair inhibitor
  • Non-small-cell lung cancer
  • NQO1

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

An NQO1- and PARP-1-mediated cell death pathway induced in non-small-cell lung cancer cells by β-lapachone. / Bey, Erik A.; Bentle, Melissa S.; Reinicke, Kathryn E.; Dong, Ying; Yang, Chin Rang; Girard, Luc; Minna, John D.; Bornmann, William G.; Gao, Jinming; Boothman, David A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, No. 28, 10.07.2007, p. 11832-11837.

Research output: Contribution to journalArticle

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AU - Yang, Chin Rang

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N2 - Lung cancer is the number one cause of cancer-related deaths in the world. Patients treated with current chemotherapies for non-small-cell lung cancers (NSCLCs) have a survival rate of ≈15% after 5 years. Novel approaches are needed to treat this disease. We show elevated NAD(P)H:quinone oxidoreductase-1 (NQO1) levels in tumors from NSCLC patients. β-Lapachone, an effective chemotherapeutic and radiosensitizing agent, selectively killed NSCLC cells that expressed high levels of NQO1. Isogenic H596 NSCLC cells that lacked or expressed NQO1 along with A549 NSCLC cells treated with or without dicoumarol, were used to elucidate the mechanism of action and optimal therapeutic window of β-lapachone. NSCLC cells were killed in an NQO1-dependent manner by β-lapachone (LD50, ≈4 μM) with a minimum 2-h exposure. Kinetically, β-lapachone-induced cell death was characterized by the following: (i) dramatic reactive oxygen species (ROS) formation, eliciting extensive DNA damage; (ii) hyperactivation of poly(ADP-ribose)polymerase-1 (PARP-1); (iii) depletion of NAD+/ATP levels; and (iv) proteolytic cleavage of p53/PARP-1, indicating μ-calpain activation and apoptosis. β-Lapachone-induced PARP-1 hyperactivation, nucleotide depletion, and apoptosis were blocked by 3-aminobenzamide, a PARP-1 inhibitor, and 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM), a Ca2+ chelator. NQO1- cells (H596, IMR-90) or dicoumarol-exposed NQO1+ A549 cells were resistant (LD50, >40 μM) to ROS formation and all cytotoxic effects of β-lapachone. Our data indicate that the most efficacious strategy using β-lapachone in chemotherapy was to deliver the drug in short pulses, greatly reducing cytotoxicity to NQO1- "normal" cells. β-Lapachone killed cells in a tumor-selective manner and is indicated for use against NQO1+ NSCLC cancers.

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