Molecular Mechanism of the Cell Death Induced by the Histone Deacetylase Pan Inhibitor LBH589 (Panobinostat) in Wilms Tumor Cells.

Tao Yan-Fang,Feng Xing,Zhao He,Liu Lin,Wu Yi,Pan Jian,Li Gang,Su Guang-Hao,Xiao Pei-Fang,Wang Jian,Ni Jian,Fang Fang,Lu Jun,Liang‐Guo Xu ,Zhiheng Li ,Lichao Sun ,Xiaohong Du ,Meifang Jin ,Yanhong Li ,Huiting Zhou ,Yiping Li ,Xiaoyan Zhu ,Shaoyan Hu ,Wei Zhao ,Nana Wang ,Liu Cao ,Yiming Xu

doi:10.1371/journal.pone.0126566

Abstract

BackgroundWilms 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.MethodsSK-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.ResultsLBH589 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.ConclusionsLBH589 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.

Highlights

Wilms tumor (WT) is an embryonic cancer of the kidney composed of blastemal, stromal and epithelial elements
LBH589 treatment inhibited the growth of SK-NEP-1 xenograft tumors in nude mice
Expression profiling, and Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Ingenuity pathway analysis (IPA) analyses identified new targets and a new “network” of genes responding to LBH589 treatment in SK-NEP-1 cells

Summary

Introduction

Wilms tumor (WT) is an embryonic cancer of the kidney composed of blastemal, stromal and epithelial elements. WT is the most common malignant neoplasm of the urinary tract in children [1]. The overall 5-year survival is estimated as > 80% [4]; for individuals, the prognosis is highly dependent on individual staging and treatment. WT is almost curable, with long-term survival, the combination of chemotherapy, radiotherapy and surgery often results in severe complications in adulthood [2]. Decreases the treatment burden and improve outcome of patients are still required [3]. We evaluated the efficacy of LBH589, a histone deacetylases (HDACs) pan inhibitor to inhibit WT development in vitro and in vivo

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Results