6533b828fe1ef96bd1288f8c

RESEARCH PRODUCT

Corrigendum to "poly (ADP-ribose) polymerase inhibition synergizes with the NF-κB inhibitor DHMEQ to kill hepatocellular carcinoma cells" [Biochim. Biophys. Acta 1853(12)(2015) 3224-3234].

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Fig. 1. The effects of the DHMEQ–Olaparib combination on HCC cells. (A) Cells were treated for 72 hwith the indicated concentrations of DHMEQ–Olaparib and cell viability was assessed by MTS assays. The DHMEQ–Olaparib combination showed synergistic inhibition of cell viability in Hep3B cells and additive inhibition in Huh7 cells. Combination index (CI) values are indicated above the bar. Data are expressed as percent cell growth and are the mean ± SD of three separate experiments (each of which was performed in triplicate). *p b 0.05 and **p b 0.01 versus each agent alone. (B) Cells were treated for 24 h with DHMEQ (μg/ml) or Olaparib (μM) alone or in combination, allowed to grow for 14 days in the absence of drugs and surviving colonies stained and counted. Data are expressed as percent of control cell colonies and are the mean ± SD of two separate experiments (each of which was performed in duplicate). *p b 0.05 versus each agent alone. (C) Cells were treated with the indicated concentrations of DHMEQ (μg/ml) or Olaparib (μM) alone or in combination for 24 h and cell proliferation was assessed by the BrdU assay. Data are expressed as percent cell proliferation and are the means ± SD of three separate experiments (each of which was performed in triplicate). **p b 0.01 versus each agent alone. The DHMEQ–Olaparib combination showed synergistic inhibition of cell proliferation in Hep3B and additive inhibition in Huh7 cells. The combination index (CI) values are indicated above the bar. (D) Cells were treated for 24 h with DHMEQ (μg/ml) or Olaparib (μM) alone or in combination, and caspase 3/7 activation was determined by caspase assays. Data are expressed as fold increase from untreated cells and are the means ± SD of two separate experiments, each of which was performed in triplicate. *p b 0.05 versus each agent alone. (E) Cellswere treatedwithDHMEQ(μg/ml) or Olaparib (μM)alone or in combination for 24 h. The induction of PARP cleavage (85 kDa) and changes in Bcl-2 expression level were analysed bywestern blotting. The numbers represent fold of Bcl-2 differencewith vehicle-treated control samples (DMSO) arbitrarily set at 1.0. The data represent two independent representative experiments. (F) Expression of Bcl2 mRNA was analysed by q-PCR in Hep3B cells. Relative expression was calculated as ratio of drug-treated samples versus control (DMSO) and corrected by the quantified expression level of β-actin. The results shown are the means ± SD of two experiments (each performed in triplicate). **p b 0.01 versus each agent alone. (Right) Expression of IL-6 mRNA was analysed by semiquantitative-PCR in Hep3B cells using gene-specific primers. The results were normalized to the expression of β-actin for all the samples. The results shown are the means ± SD of two experiments. *p b 0.05 versus each agent alone. (G) Hep3B cells were treated for 24 h with DHMEQ (μg/ml) and Olaparib (μM) either alone or in combination and apoptotic cells were visualized by TUNEL staining as described in the Materials and methods section. (Right) Bar charts show the percentage of DHMEQ–Olaparib-induced apoptotic cells. Data are expressed as the means ± SD of two separate experiments. Magnification 20×. Scale bar = 50 μM. **p b 0.01 versus each agent alone. (H) Hep3B cells were treated for 24 h with DHMEQ (μg/ml) or Olaparib (μM) alone or in combination and mRNA expression levels of HKR were assessed by RT-PCR semi-quantitative analysis. The data represent two independent experiments with comparable outcomes. (I) Hep3B cells were transfected with a Non-Correlated siRNA (siRNA NC) or Harakiri siRNA (siRNA HKR). (Right) RT-PCR semi-quantitative analysis of HKR mRNA in cells transfected. 48 h after transfection. (Left) Cells were treated for 24 h with DHMEQ (μg/ ml) orOlaparib (μM)alone or in combination and cell viabilitywas assessed by theMTS assays. The data aremean±SDof three separate experiments each ofwhichwas performed in triplicate. *p b 0.05 versus siRNA NC.