title: Effects of JS-K, a novel anti-cancer nitric oxide prodrug, on gene expression in human hepatoma Hep3B cells.
abstract: JS-K is a novel anticancer nitric oxide (NO) prodrug effective against a variety of cancer cells, including the inhibition of AM-1 hepatoma cell growth in rats. To further evaluate anticancer effects of JS-K, human hepatoma Hep3B cells were treated with JS-K and the compound control JS-43-126 at various concentrations (0-100μM) for 24h, and cytotoxicity was determined by the MTS assay. The compound control JS-43-126 was not cytotoxic to Hep3B cells at concentrations up to 100μM, while the LC50for JS-K was about 10μM. To examine the molecular mechanisms of antitumor effects of JS-K, Hep3B cells were treated with 1-10μM of JS-K for 24h, and then subjected to gene expression analysis via real time RT-PCR and protein immunostain via confocal images. JS-K is a GST-α targeting NO prodrug, and decreased immunostaining for GST-α was associated with JS-K treatment. JS-K activated apoptosis pathways in Hep3B cells, including induction of caspase-3, caspase-9, Bax, TNF-α, and IL-1β, and immunostaining for caspase-3 was intensified. The expressions of thrombospondin-1 (TSP-1) and the tissue inhibitors of metalloproteinase-1 (TIMP-1) were increased by JS-K at both transcript and protein levels. JS-K treatment also increased the expression of differentiation-related genes CD14 and CD11b, and depressed the expression of c-myc in Hep3B cells. Thus, multiple molecular events appear to be associated with anticancer effects of JS-K in human hepatoma Hep3B cells, including activation of genes related to apoptosis and induction of genes involved in antiangiogenesis and tumor cell migration.
authors.affiliation: University of Southern California, Los Angeles, CA, United States. Electronic address: raydong510@gmail.com.
authors.forename: Ray
authors.initials: R
authors.lastname: Dong
authors.affiliation: The Ohio State University, Columbus, OH, United States.
authors.forename: Xueqian
authors.initials: X
authors.lastname: Wang
authors.affiliation: Zunyi Medical College, Zunyi, China. Electronic address: wh@zmc.edu.cn.
authors.forename: Huan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Zunyi Medical College, Zunyi, China.
authors.forename: Zhengyun
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Zunyi Medical College, Zunyi, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Leidos Biomedical Research Inc, Frederick, MD, United States.
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Saavedra
mesh_terms: D000209:Acute-Phase Proteins / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D001391:Azo Compounds / Q000737:chemistry / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000098:blood supply / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008113:Liver Neoplasms / Q000098:blood supply / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D020782:Matrix Metalloproteinases / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000473:pathology; D009569:Nitric Oxide / Q000378:metabolism*; D010879:Piperazines / Q000737:chemistry / Q000494:pharmacology; D011355:Prodrugs / Q000494:pharmacology*; D019699:Thrombospondins / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism
chemical_list: D000209:Acute-Phase Proteins; D000970:Antineoplastic Agents; D001391:Azo Compounds; D007527:Isoenzymes; C474958:O(2)-(2,4-dinitrophenyl) 1-((4-ethoxycarbonyl)piperazin-1-yl)diazen-1-ium-1,2-diolate; D010879:Piperazines; D011355:Prodrugs; D019699:Thrombospondins; D019715:Tissue Inhibitor of Metalloproteinase-1; D009569:Nitric Oxide; D005982:Glutathione Transferase; C119995:glutathione S-transferase alpha; D053148:Caspase 3; D020782:Matrix Metalloproteinases
keywords: Apoptosis; Differentiation; Gene expression; Hep3B cells; JS-K; Nitric oxide donor