Published: March 01, 2011 r2011 American Chemical Society 2284 dx.doi.org/10.1021/jf103488j | J. Agric. Food Chem. 2011, 59, 2284–2290 ARTICLE pubs.acs.org/JAFC (-)-Anonaine Induces DNA Damage and Inhibits Growth and Migration of Human Lung Carcinoma H1299 Cells Bing-Hung Chen, † Hsueh-Wei Chang, ‡ Hsuan-Min Huang, ‡ Inn-Wen Chong, § Jia-Shing Chen, || Chung-Yi Chen,* ,^ and Hui-Min Wang* ,# † Department of Biotechnology, Kaohsiung Medical University, Kaohsiung 807, Taiwan ‡ Department of Biomedical Science and Environmental Biology, Graduate Institute of Natural Products, College of Pharmacy, Center of Excellence for Environmental Medicine, Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung 807, Taiwan § Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan ) Institute of Molecular Medicine, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan ^ 151, Ching-Hsueh Road, Ta-Liao District, Department of Medical Laboratory Science and Biotechnology, School of Medical and Health Sciences, Fooyin University, Kaohsiung County 831, Taiwan, R.O.C. # 100, Shih-Chuan 1st Road, San-Ming District, Department of Fragrance and Cosmetic Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan, R.O.C. ABSTRACT: The anticancer effects of (-)-anonaine were investigated in this current study. (-)-Anonaine at concentration ranges of 50-200 μM exhibited significant inhibition to cell growth and migration activities on human lung cancer H1299 cells at 24 h, albeit cell cycle analyses showed that (-)-anonaine at the above concentration ranges did not cause any significant changes in cell- cycle distributions. Significant nuclear damages of H1299 cells were observed with 10-200 μM(-)-anonaine treatment in a comet assay, whereas higher concentrations (6 and 30 mM) of (-)-anonaine concentrations were required to cause DNA damages in an in vitro plasmid cleavage assay. In summary, our results demonstrated that (-)-anonaine exhibited dose-dependent antiproliferatory, antimigratory, and DNA-damaging effects on H1299 cells. We inferred that (-)-anonaine can cause cell-cycle arrest and DNA damage to hamper the physiological behavior of cancer cells at 72 h, and therefore, it can be useful as one of the potential herbal supplements for chemoprevention of human lung cancer. KEYWORDS: (-)-Anonaine, lung cancer, antiproliferation, DNA damage, comet assay, cell migration ’ INTRODUCTION Lung cancer is one of the leading deaths in both males and females globally. 1 Non-small cell lung cancer (NSCLC) is lung cancer of any epithelial type (including adenocarcinoma, squamous cell carcinoma, and large cell carcinoma) other than small cell lung cancer (SCLC). NSCLCs account for approximate 85% of all lung cancer cases and are relatively insensitive to chemotherapy clinically, as compared to SCLCs. 2 Although various strategies for treating lung cancers, such as chemotherapy, radiotherapy, and radiosurgery, have been reported, 3 development of novel and effective drugs against lung cancers still remains a challenge. Michelia alba DC (Magnoliaceae) is a flowering plant native to tropical and subtropical southeast Asia, whose various constituents have been widely used for medicinal purposes. While extracts from its white yellowish fragrant flowers are a potent smooth-muscle relaxant and used as an abortive agent, the bark of M. alba DC is commonly used to treat malaria, syphilis, and gonorrhea in oriental medicine. 4 (-)-Anonaine (Figure 1), an aporphine isoquinoline alkaloid isolated from leaves of M. alba DC, has been shown to exhibit various phar- macological effects that include antidepressant, 5 antiperoxida- tive, 6 antifungal, and antibacterial activities. 7 However, the antitumor activities of (-)-anonaine are poorly understood and remain to be elucidated. In our current work, we investigated the anticancer potential of (-)-anonaine by measuring its cytotoxic, DNA-damaging, and antimigratory effects on human lung carcinoma H1299 cells. Effects of (-)-anonaine on cell-cycle progression were also examined. Our data suggested that (-)-anonaine could be beneficial in inhibiting both growth and migration of lung cancers, and its underlying mechanisms of such anticancer activities merit further investigation. This is the first study to demonstrate (-)- anonaine biofunctions on H1299 human lung cancer cells. ’ MATERIALS AND METHODS Purification of (-)-Anonaine. Stems of M. alba DC were collected from Fooyin University, Kaohsiung County, Taiwan, in March 2005. A voucher specimen (Michelia 2) was characterized by Dr. Yen- Ray Hsui of the Division of Silviculture, Taiwan Forestry Research Institute, Taipei, Taiwan, and deposited in the School of Medical and Health Sciences, Fooyin University, Kaohsiung County, Taiwan. The air-dried stems of M. alba DC (3.0 kg) were extracted with methanol (MeOH) (50 L  6) at room temperature, and a MeOH extract (156.7 Received: September 9, 2010 Accepted: January 6, 2011 Revised: January 6, 2011