Journal: Journal of Virology

Article Title: Cellular DNA Topoisomerases Are Required for the Synthesis of Hepatitis B Virus Covalently Closed Circular DNA

doi: 10.1128/JVI.02230-18

Figure Lengend Snippet: Effects of topoisomerase poisons on cccDNA synthesis

Article Snippet: Topotecan (HY-13768A) and idarubicin (HY-17381) were purchased from MedChemExpress (MCE).

Techniques: Control

Journal: Journal of Virology

Article Title: Cellular DNA Topoisomerases Are Required for the Synthesis of Hepatitis B Virus Covalently Closed Circular DNA

doi: 10.1128/JVI.02230-18

Figure Lengend Snippet: TOP1 and TOP2 inhibitors block cccDNA synthesis. (A) Schematic presentation of the experimental schedule. HepAD38 cells were cultured in the absence of Tet, and 2 mM PFA was added to the culture medium 2 days after Tet removal to arrest viral DNA synthesis. Four days later, while PFA was withdrawn, Tet was added back to the culture medium to stop viral pgRNA transcription from the transgene. Cells were left untreated or treated with topotecan (TPT; 1 μM) or doxorubicin (Doxo; 1 μM) at 16 h after PFA removal and harvested at the indicated time points. (B) Hirt DNA after heat denaturalization at 88°C for 8 min and EcoRI digestion was resolved by agarose gel electrophoresis and HBV DNA species were detected by Southern blot hybridization with a riboprobe specifically hybridizing to negative-strand DNA. mtDNA served as a loading control. (C) The amounts of cccDNA were quantified by phosphorimager, normalized to the amount of mtDNA, and plotted as the percentage of that in the mock-treated (UT) cells harvested at 16 h post-PFA removal. Means and standard deviations (n = 4) are presented. *DP-rc, denatured deproteinized rc DNA; ccc*, EcoRI-linearized cccDNA.

Article Snippet: Topotecan (HY-13768A) and idarubicin (HY-17381) were purchased from MedChemExpress (MCE).

Techniques: Blocking Assay, Cell Culture, DNA Synthesis, Agarose Gel Electrophoresis, Southern Blot, Hybridization, Control

Journal: Journal of Virology

Article Title: Cellular DNA Topoisomerases Are Required for the Synthesis of Hepatitis B Virus Covalently Closed Circular DNA

doi: 10.1128/JVI.02230-18

Figure Lengend Snippet: TOP1 and TOP2 inhibitors inhibited HBV cccDNA synthesis in de novo infection. (A) Schematic representation of the experimental schedule. C3AhNTCP cells were infected with HBV at an MOI of 250 genome equivalents for 24 h. The cells were mock treated or treated with 200 nM doxorubicin (Doxo), 200 nM topotecan (TPT), 200 nM doxorubicin and 200 nM topotecan (Doxo + TPT), 200 nM aclarubicin (Acla), or 1 μg/ml myrcludex B (Myr-B) starting from HBV infection for a total of 36 h. (B) Hirt DNA was resolved by agarose gel electrophoresis after heat denaturalization at 88°C for 8 min and EcoRI digestion. HBV DNA species were detected by Southern blot hybridization with a riboprobe specifically hybridizing to negative-strand DNA. mtDNA served as a loading control of Hirt DNA analysis. (C) cccDNAs were quantified by a phosphorimager. The data from three independent experiments are presented. P values calculated by Student's t test are presented.

Article Snippet: Topotecan (HY-13768A) and idarubicin (HY-17381) were purchased from MedChemExpress (MCE).

Techniques: Infection, Agarose Gel Electrophoresis, Southern Blot, Hybridization, Control

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: IC50 Values for small molecule inhibitors and topotecan in neuroblastoma cell lines

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques:

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: A. Shows a representative bar graph for the combination effects of inhibitors and topotecan on neuroblastoma cells growth treated with fixed one concentration of each inhibitor at sub-IC50 concentration for 72 hours. For combination treatments, SH-SY-5Y cells were treated with 13-197, BI2536, vismodegib and topotecan at 5 μM, 5 nM, 50 μM, and 5 nM concentrations, respectively; IMR-32 cells were treated with 13-197, BI2536, vismodegib and topotecan at 5 μM, 5 nM, 50 μM, and 10 nM concentrations, respectively; and SK-N-BE(2) were treated with 13-197, BI2536, vismodegib and topotecan at 5 μM, 5 nM, 50 μM and 50 nM concentrations, respectively. Following treatment, the cells were subjected to growth analyses using MTT assay. B-D. MTT assay showing the combination effects of indicated small molecule inhibitors and topotecan on neuroblastoma cells growth at 72 hours in a dose-dependent manner. The values represent the means ± SD from four wells of 96-well plates. *, p<0.05.

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques: Concentration Assay, MTT Assay

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: Combination indexes of small molecule inhibitors and topotecan in neuroblastoma cell lines

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques:

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: A. Illustrates a representative scatter diagram for the apoptotic cell analyses following treatment as indicated above in Figure with small molecule inhibitors alone or in combination with topotecan in neuroblastoma cells. B. Quantification of the apoptotic cells (% Annexin-V/PI double positive) following small molecule inhibitors alone or in combination with topotecan treatment in SH-SY-5Y, IMR-32 and SK-N-BE(2) neuroblastoma cells. The values represent the means ± SD of three separate experiments. *, p<0.05.

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques:

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: Each neuroblastoma cell line was treated with inhibitors alone or combination with topotecan (as described in Figure ) for 24 hours and the expression of associated pathways/molecules were determined using western blot analyses. β-Actin was used as a loading control in these experiments.

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques: Expressing, Western Blot, Control

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: A. Shows a representative micrograph of small, medium and large neurospheres in 13-197 (5 μM), BI2536 (5 nM) and vismodegib (50 μM) alone or in combination with topotecan (10 nM) treated IMR-32cells. The micrographs were taken using a phase contrast microscope at 4X magnification. Scale bar; 100 μm. B. Quantification of sphere assay data in above described treated cells. The measurement of 50 to <100 μm size for small, >100 to 200 μm for medium and >200 μm size for the large neurosphere were considered. The values represent the means ± SD from three wells of 6-well plates. *, p<0.01. C. Western blot analyses for the Nestin and CD133 expression following the indicated treatments of neurospheres. β-Actin was used as a loading control in this experiment.

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques: Microscopy, Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Improved therapy for neuroblastoma using a combination approach: superior efficacy with vismodegib and topotecan

doi: 10.18632/oncotarget.7714

Figure Lengend Snippet: NSG mice bearing tumors were treated with vehicle (DMSO) or vismodegib (50 mg/kg) and topotecan (10 mg/kg) alone, or both, twice a week for four weeks. For the combination, mice were treated with each agent first (denoted as Pre-) and then next day with the other agent. A. Tumor growth analyses following treatments. B. Kaplan–Meier analyses for the survival of mice using the log-rank test. C. Histological (H&E) and immunohistochemical (Smo, Ki-67, MYCN and cleaved caspase3) analyses of tumors in mice treated as indicated. The images were scanned and captured using a digital scanner VENTANA Image software (Roche, Germany) at ×40 magnification.

Article Snippet: Hedgehog inhibitor vismodegib was purchased from LC laboratories (Woburn, MA) and PLK1 inhibitor BI2536 and chemotherapy topotecan were purchased from Selleckchem Company (Houston TX).

Techniques: Immunohistochemical staining, Software

Journal: NPJ Precision Oncology

Article Title: Ex vivo 3D micro-tumour testing platform for predicting clinical response to platinum-based therapy in patients with high-grade serous ovarian cancer

doi: 10.1038/s41698-025-01080-8

Figure Lengend Snippet: A Heatmap showing ex vivo patient responses to first and second-line treatments. Colour represents AUC normalized per treatment. B Ex vivo dose response curves of micro-tumours treated with carboplatin, paclitaxel, gemcitabine, topotecan and doxorubicin, respectively. Example patients for patient report (see Fig. 5C) are highlighted. Points represent mean and standard deviation of technical replicates. C Responses to measured treatment options for a specific patient relative to the full cohort of patients. Each horizontal green-to-blue bar indicates the range of AUCs measured for a particular drug in the entire cohort of patients. The vertical line indicates the patient’s percentile within this range.

Article Snippet: The following drugs were included: carboplatin (Fresenius Kabi, Cat# 08717371358198, 16 concentrations in technical duplicates), paclitaxel (Selleckchem, Cat# s1150, 16 concentrations in technical duplicates), gemcitabine (Selleckchem, Cat# s1149, 16 concentrations in technical duplicates), doxorubicin (Selleckchem, Cat# s1208), topotecan (Selleckchem, Cat# s1231, 12–16 concentrations in technical duplicates), olaparib (Selleckchem, Cat# s1060, 12–16 concentrations in technical duplicates) and niraparib (MedChemExpress, Cat# HY-10619, 12–16 concentrations in technical duplicates).

Techniques: Ex Vivo, Standard Deviation

Journal: Molecular Psychiatry

Article Title: Frontotemporal dementia patient-derived iPSC neurons show cell pathological hallmarks and evidence for synaptic dysfunction and DNA damage

doi: 10.1038/s41380-025-03272-x

Figure Lengend Snippet: NUPR2 , potentially related to the DNA damage pathway, is the only differentially expressed gene when comparing the C9-HRE and sporadic FTD neurons ( A ). Both sporadic (C9-) and C9-HRE-carrying FTD (C9+) neurons display nuclei, which are significantly rounder in shape as well as significantly smaller compared to healthy neuron nuclei, indicated by altered nuclear eccentricity ( B ), and a higher number of micronuclei ( C ). A representative image of a micronucleus next to the nucleus is shown (arrow). Kruskal-Wallis test followed by Dunn’s multiple comparisons test was used. n [Control] = 74; n [C9−] = 17; n [C9+] = 21. In addition, the distribution of larger and smaller sized nuclei is different in FTD neurons than that in control neurons ( D – F ). Representative images of control, C9-, and C9+ neurons treated with vehicle (upper panel) or 10 µM topotecan (lower panel). The neurons were stained with antibodies against MAP2 (red) and γH2A.X (green). Nuclei were stained with DAPI (blue) ( G ). Topotecan treatment significantly increases the number of γH2A.X-positive foci in the nuclei in all neurons, indicating increased DNA damage. At baseline, C9+ neurons display a slightly higher number of γH2A.X foci compared to control neurons, although this difference is not statistically significant ( H – J ). Kruskal-Wallis test followed by Dunn’s multiple comparisons test was used. n [Control] = 26–28; n [C9−] = 15–18; n [C9+] = 44–64. Statistically significant differences are shown as *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: To study the DNA damage response, iPSC-derived neurons were treated with 10 μM topotecan (Topotecan hydrochloride, Tocris, 4562) or DMSO vehicle for 1 h. The cells were fixed with 4% PFA and stained with conjugated MAP2 and γH2A.X antibody as described above.

Techniques: Control, Staining

Journal: Cancer science

Article Title: Histone deacetylase inhibitors enhance the chemosensitivity of tumor cells with cross-resistance to a wide range of DNA-damaging drugs.

doi: 10.1111/j.1349-7006.2007.00669.x

Figure Lengend Snippet: Fig. 5. HC-toxin selectively enhances the induction of cell death by a variety of DNA-damaging agents. (a) OVCAR-8 or SKOV-3 cells were incubated for 48 h with the indicated concentrations of bleomycin (BLM), mitomycin-C (MMC), topotecan (TPT), etoposide (VP-16), doxorubicin (DXR), 5-fluorouracil (5-FU), vincristine (VCR), or paclitaxel (PTX), in the absence (–HC) or presence (+HC) of 0.1 µM HC-toxin. The proportion of cells in sub-G1 phase was then determined using flow cytometry. Data are means ± SD from three separate experiments, each performed in duplicate. (b) OVCAR-8 cells were incubated for 9 h with 50 µM BLM, 2 µM MMC, 0.5 µM TPT, 20 µM VP-16, or 0.5 µM DXR, in the absence or presence of 0.1 µM HC-toxin. They were then fixed and subjected to immunofluorescence analysis with antibodies to phosphorylated H2AX. The cells were counterstained with antibodies to β-actin. Bar = 100 µm. Data are representative of three separate experiments. (c) OVCAR-8 cells were incubated for 24 h (upper panel) or 48 h (lower panel) as in (b) but in the additional absence or presence of 10 mM NAC, as indicated. The proportion of cells manifesting ROS accumulation (upper panel) or a fractional DNA content (lower panel) was then determined using flow cytometry. Data are means ± SD from three separate experiments, each performed in duplicate. *P < 0.01, **P < 0.001.

Article Snippet: Mitomycin-C was from ICN Biochemicals (Irvine, CA, USA), topotecan was from LKT Laboratories (St Paul, MN, USA), MS-275 was from Calbiochem (La Jolla, CA, USA), and FK228 was kindly provided by Fujisawa Pharmaceutical Co. (Osaka, Japan).

Techniques: Incubation, Flow Cytometry, Immunofluorescence

Journal: Biomarker Research

Article Title: Combination statin and chemotherapy inhibits proliferation and cytotoxicity of an aggressive natural killer cell leukemia

doi: 10.1186/s40364-018-0140-0

Figure Lengend Snippet: Combination of statins and chemotherapy augment the inhibition of proliferation and cytotoxicity. YT-INDY cells were incubated with topotecan, paclitaxel or doxorubicin in the presence or absence of statins for 72 h. Cell growth and cellular cytotoxicity was measured and expressed as a percent of cell control, which was treated with the solvent in which the drug was dissolved. a Addition of atorvastatin, fluvastatin or simvastatin to the chemotherapy drugs enhanced the inhibition of cell growth compared to either drug alone at a statistical significance level of p < 0.05 when compared to the single drug controls. b Addition of statins to paclitaxel-treated YT-INDY demonstrated an enhance inhibition of cellular cytotoxicity, which was statistically significant at the p < 0.05 level for all statin/paclitaxel combinations. A similar effect was not observed with topotecan or doxorubicin. Control levels of YT-INDY cytotoxicity against the target cell line averaged 61.6% ± 3.32 (standard error of the mean) ( n = 16 experiments)

Article Snippet: The chemotherapeutic agents used included doxorubicin (Fisher Scientific, Waltham, MA, USA), paclitaxel (LKT Laboratories Inc.. St. Paul, MN, USA) and topotecan (Toronto Research Chemicals, Toronto, Canada).

Techniques: Inhibition, Incubation, Control, Solvent