title: Nuciferine, extracted from Nelumbo nucifera Gaertn, inhibits tumor-promoting effect of nicotine involving Wnt/β-catenin signaling in non-small cell lung cancer.
abstract: BACKGROUND
  The leaves of Nelumbo nucifera Gaertn are recorded in the earliest written documentation of traditional Chinese medicinal as "Ben Cao Gang Mu", a medicinal herb for blood clotting, dysentery and dizziness. Recently, nuciferine (NF), one of N. nucifera Gaertn leaf extracts has been shown to possess several pharmacological properties, including anti-viral and anti-cancer. The aim of this study is to investigate the underlying molecular mechanism of the anti-cancer activity of NF in NSCLC progression induced by nicotine
  
  
  METHODS
  The effect of NF on proliferation of A549 (human lung adenocarcinoma epithelial cell line) pretreated with or without nicotine was detected by tumor cell proliferation assay. TOP-Flash reporter assay was applied to investigate the activity of Wnt/β-catenin signaling in tumor cells in the presence of NF and/or nicotine. Apoptosis was measured using a FITC-Annexin V and PI detection kit by flow cytometry. In addition, mRNA or protein expression levels were respectively tested by quantitative RT-PCR or western blot. In vivo experiments, tumor samples were fixed in formalin and embedded in paraffin for additional analyses by immunohistochemistry and TUNEL staining.
  
  
  RESULTS
  NF significantly inhibited the proliferation of NSCLC cells in the presence of nicotine, suppressed the activity of Wnt/β-catenin signaling, enhanced the stabilization of Axin, and induced apoptosis. NF down-regulated the expression levels of β-catenin and its downstream targets including c-myc, cyclin D and VEGF-A. NF also decreased the ratio of Bcl-2/Bax, which may explain the pro-apoptosis effect of NF. In tumor xenograft nude mice, NF not only inhibited the growth of non-small cell lung cancer (NSCLC) cells, but also remarkably alleviated the injury induced by nicotine in liver function.
  
  
  CONCLUSIONS
  NF has the remarkable effect to inhibit nicotine-induced NSCLC progression, which was due to its ability to reduce the activity of Wnt/β-catenin signaling. Thus, the work stated here emphasizes the importance of this traditional medicine and presents a potential novel alternative to NSCLC prevention and therapy.
authors.affiliation: Department of Public Health, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Road, Wuhan 430030, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, No. 13 Hangkong Road, Wuhan 430030, China.
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Yi
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, No. 13 Hangkong Road, Wuhan 430030, China.
authors.forename: Jian-Li
authors.initials: JL
authors.lastname: Guo
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, No. 13 Hangkong Road, Wuhan 430030, China.
authors.forename: Zhu-Xing
authors.initials: ZX
authors.lastname: Xiang
authors.affiliation: Department of Blood Transfusion, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 JieFang Road, Wuhan 430030, China.
authors.forename: Lin-Feng
authors.initials: LF
authors.lastname: Deng
authors.affiliation: Department of Blood Transfusion, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 JieFang Road, Wuhan 430030, China. Electronic address: 248961279@qq.com.
authors.forename: Lei
authors.initials: L
authors.lastname: He
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001060:Aporphines; D002289:Carcinoma, Non-Small-Cell Lung / Q000139:chemically induced / Q000188:drug therapy*; D045744:Cell Line, Tumor; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use*; D006801:Humans; D008175:Lung Neoplasms / Q000139:chemically induced / Q000188:drug therapy*; D008297:Male; D008516:Medicine, Chinese Traditional / Q000379:methods; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D031653:Nelumbo / Q000737:chemistry*; D009368:Neoplasm Transplantation; D009538:Nicotine / Q000037:antagonists & inhibitors* / Q000633:toxicity; D018515:Plant Leaves / Q000737:chemistry*; D060449:Wnt Signaling Pathway / Q000187:drug effects* / Q000502:physiology
chemical_list: D001060:Aporphines; D004365:Drugs, Chinese Herbal; D009538:Nicotine; C008692:nuciferine
keywords: Nicotine; Non-small cell cancer; Nuciferine; Proliferation; Wnt/β-catenin