Nicotine prevents the apoptosis induced by menadione in human lung cancer cells

https://doi.org/10.1016/j.bbrc.2006.02.047Get rights and content

Abstract

Approximately 50% of long-term cigarette smokers die prematurely from the adverse effects of smoking, including on lung cancer and other illnesses. Nicotine is a main component in tobacco and has been implicated as a potential factor in the pathogenesis of human lung cancer. However, the mechanism of nicotine action in the development of lung cancer remains largely unknown. In the present study, we designed a nicotine-apoptosis system, by pre-treatment of nicotine making lung cancer cell A549 to be in a physiological nicotine environment, and observed that nicotine promoted cell proliferation and prevented the menadione-induced apoptosis, and exerts its role of anti-apoptosis by shift of apoptotic stage induced by menadione from late apoptotic stage to early apoptotic stage, in which NF-κB was up-regulated. Interference analysis of NF-κB in A549 cells showed that knock down of NF-κB resulted in apoptosis promotion and counteracted the protective effect of nicotine. The findings suggest that nicotine has potential effect in lung cancer genesis, especially in patients with undetectable early tumor development and development of specific NF-κB inhibitors would represent a potentially exciting new pharmacotherapy for tobacco-related lung cancer.

Section snippets

Materials and methods

Cell culture. Human lung adenocarcinoma cell line A549 was obtained from Centre for Type Culture Collection (Wuhan, China). The cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum at 37 °C, with 5% CO2. Medium was replaced every other day.

MTT assay. Cell viability was determined using the MTT assay kit according to the manufacturer’s protocol (Amresco, Solon, OH, USA). After the treatments with nicotine hydrogen tartrate salt (Sigma, St. Louis, MO,

Nicotine promoted cell proliferation and prevented the apoptosis

To study roles of nicotine in apoptosis, menadione was first used to set up apoptosis system in lung cancer cells A549. The cells were treated with concentration gradient of menadione as 1–50–100–200 μM. The apoptotic cells turned round and their nucleus broke into pieces, forming apoptotic bodies (Figs. 1A–D). Apoptotic cells were obviously observed at concentrations of 100 μM menadione by FITC-labeled TdT-mediated dUTP nick end labeling (TUNEL) assay (Figs. 1E–H). Quantitative analysis using

Discussion

Both hereditary and environmental factors contributed to the genesis of lung cancer, which involved many genes including those in processes of DNA reparation, signal transduction, and cell cycle regulation [10]. Defects of these processes will result in activation of oncogenes, the inactivation of anticancer genes, and genomic unstabilization, and thus lung cancer genesis [11], [12], [13]. Although some factors were elucidated to play a role in lung cancer genesis, the molecular mechanisms are

Acknowledgments

The work was supported by the National Natural Science Foundation of China, the National Key Basic Research project (G2000057004), the Program for New Century Excellent Talents in University and the Key Project of Chinese Ministry of Education (No. 2004.28). There is not any financial conflict of interest.

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