title: Molecular Mechanism of the Cell Death Induced by the Histone Deacetylase Pan Inhibitor LBH589 (Panobinostat) in Wilms Tumor Cells.
abstract: BACKGROUND
  Wilms tumor (WT) is an embryonic kidney cancer, for which histone acetylation might be a therapeutic target. LBH589, a novel targeted agent, suppresses histone deacetylases in many tumors. This study investigated the antitumor activity of LBH589 in SK-NEP-1 and G401 cells.
  
  
  METHODS
  SK-NEP-1 and G401 cell growth was assessed by CCK-8 and in nude mice experiments. Annexin V/propidium iodide staining followed by flow cytometry detected apoptosis in cell culture. Gene expressions of LBH589-treated tumor cells were analyzed using an Arraystar Human LncRNA Array. The Multi Experiment View cluster software analyzed the expression data. Differentially expressed genes from the cluster analyses were imported into the Ingenuity Pathway Analysis tool.
  
  
  RESULTS
  LBH589 inhibited cell proliferation of SK-NEP-1 and G401 cells in a dose-dependent manner. Annexin V, TUNEL and Hochest 33342 staining analysis showed that LBH589-treated cells showed more apoptotic features compared with the control. LBH589 treatment inhibited the growth of SK-NEP-1 xenograft tumors in nude mice. Arraystar Human LncRNA Array analysis of genes and lncRNAs regulated by LBH589 identified 6653 mRNAs and 8135 lncRNAs in LBH589-treated SK-NEP-1 cells. The most enriched gene ontology terms were those involved in nucleosome assembly. KEGG pathway analysis identified cell cycle proteins, including CCNA2, CCNB2, CCND1, CCND2, CDK4, CDKN1B and HDAC2, etc. Ingenuity Pathway Analysis identified important upstream molecules: HIST2H3C, HIST1H4A, HIST1A, HIST1C, HIST1D, histone H1, histone H3, RPRM, HSP70 and MYC.
  
  
  CONCLUSIONS
  LBH589 treatment caused apoptosis and inhibition of cell proliferation of SK-NEP-1and G401 cells. LBH589 had a significant effect and few side effects on SK-NEP-1 xenograft tumors. Expression profiling, and GO, KEGG and IPA analyses identified new targets and a new "network" of genes responding to LBH589 treatment in SK-NEP-1 cells. RPRM, HSP70 and MYC may be important regulators during LBH589 treatment. Our results provide new clues to the proapoptotic mechanism of LBH589.
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yan-Fang
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhi-Heng
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Li-Xiao
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Fang
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Jun
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gang
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Cao
authors.initials: C
authors.lastname: Lan
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Na-Na
authors.affiliation: Department of Gastroenterology, the 5th Hospital of Chinese PLA, Yin chuan, China.
authors.forename: Du
authors.initials: D
authors.lastname: Xiao-Juan
authors.affiliation: Department of Cell and Molecular Biology, Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
authors.forename: Sun
authors.initials: S
authors.lastname: Li-Chao
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Wen-Li
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Pei-Fang
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: He
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Su
authors.initials: S
authors.lastname: Guang-Hao
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yan-Hong
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yi-Ping
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Yun-Yun
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Hui-Ting
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Wu
authors.initials: W
authors.lastname: Yi
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Mei-Fang
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Lin
authors.affiliation: Translational Research Center, Second Hospital, The Second Clinical School, Nanjing Medical University, Nanjing, China.
authors.forename: Ni
authors.initials: N
authors.lastname: Jian
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Hu
authors.initials: H
authors.lastname: Shao-Yan
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Xue-Ming
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xing
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Jian
authors.affiliation: Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Jian
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D063990:Gene Ontology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology*; D051379:Mice; D008819:Mice, Nude; D000077767:Panobinostat; D062085:RNA, Long Noncoding / Q000235:genetics; D009396:Wilms Tumor / Q000473:pathology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D007211:Indoles; D062085:RNA, Long Noncoding; D000077767:Panobinostat