title: The induction of orphan nuclear receptor Nur77 expression by n-butylenephthalide as pharmaceuticals on hepatocellular carcinoma cell therapy.
abstract: N-butylidenephthalide (BP), isolated from the chloroform extract of Angelica sinensis, has been examined for its antitumor effects on glioblastoma multiforme brain tumors; however, little is known about its antitumor effects on hepatocellular carcinoma cells. Two hepatocellular carcinoma cell lines, HepG2 and J5, were treated with either N-butylidenephthalide or a vehicle, and cell viability and apoptosis were evaluated. Apoptosis-related mRNA and proteins expressed, including orphan receptor family Nurr1, NOR-1, and Nur77, were evaluated as well as the effect of N-butylidenephthalide in an in vivo xenograft model. N-butylidenephthalide caused growth inhibition of both the cell lines at 25 microg/ml. Furthermore, N-butylidenephthalide-induced apoptosis seems to be related to Nur77 translocation from nucleus to cytosol, which leads to cytochrome c release and caspase-3-dependent apoptosis. N-butylidenephthalide-related tumor apoptosis was associated with phosphatidylinositol 3-kinase/protein kinase B (AKT)/glycogen synthase kinase-3beta rather than the mitogen-activated protein kinase or protein kinase C pathway. Blockade of AKT activation enhanced proliferation inhibition and the induction of phosphor-Bcl-2 and Nur77 proteins. Besides, the increasing apoptosis by BP via transfection wild-type cAMP-response element-binding protein (CREB) into tumor cell was suppressed by dominant phosphorylation site mutation of CREB. This finding suggested CREB pathway was also partly involved in tumor apoptosis caused by BP. Administration of N-butylidenephthalide showed similar antitumoral effects in both HepG2 and J5 xenograft tumors. N-Butylidenephthalide induced apoptosis in hepatocellular carcinoma cells, both in vitro and in vivo, suggesting a potential clinical use of this compound for improving the prognosis of hepatocellular carcinoma cells.
authors.affiliation: Graduate Institute of Biotechnology and Department of Applied Animal Science, National Ilan University, Ilan, Taiwan, Republic of China.
authors.forename: Yi-Lin
authors.initials: YL
authors.lastname: Chen
authors.forename: Min-Hui
authors.initials: MH
authors.lastname: Jian
authors.forename: Chai-Ching
authors.initials: CC
authors.lastname: Lin
authors.forename: Jung-Cheng
authors.initials: JC
authors.lastname: Kang
authors.forename: Shee-Ping
authors.initials: SP
authors.lastname: Chen
authors.forename: Po-Cheng
authors.initials: PC
authors.lastname: Lin
authors.forename: Putzer-Joseph
authors.initials: PJ
authors.lastname: Hung
authors.forename: Jen-Ren
authors.initials: JR
authors.lastname: Chen
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Chang
authors.forename: Shinn-Zong
authors.initials: SZ
authors.lastname: Lin
authors.forename: Horng-Jyh
authors.initials: HJ
authors.lastname: Harn
mesh_terms: D029971:Angelica sinensis / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy*; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D010796:Phthalic Anhydrides / Q000494:pharmacology*; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D004365:Drugs, Chinese Herbal; C539821:NR4A1 protein, human; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D010796:Phthalic Anhydrides; D011987:Receptors, Steroid; C044613:angelicae sinensis extract; C026105:butylidenephthalide