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Cancers 2018, 10, x FOR PEER REVIEW 3 of 15 (AdGFP). At 72 h post infection, in both cell lines, crystal violet staining revealed that an Cancers 2018, 10, 144 3 of 15 OAdmCherry-mediated CPE increased in a virus-dose-dependent manner. In contrast, AdGFP- treated cells did not induce CPE, even at the highest multiplicity of infection (MOI) concentration of (Figure 1A). Sensitivity to OAdmCherry was greater in MDA-MB-231 cells as compared to HCC1937 20 (Figure 1A). Sensitivity to OAdmCherry was greater in MDA-MB-231 cells as compared to cells. For example, OAdmCherry at a MOI concentration of 10 induced 50% cell viability in HCC1937 cells. For example, OAdmCherry at a MOI concentration of 10 induced 50% cell viability MDA-MB-231, whereas 74% cell viability was observed in HCC1937 (Figure 1B). Next, TMZ-mediated in MDA-MB-231, whereas 74% cell viability was observed in HCC1937 (Figure 1B). Next, TMZ- cytotoxicity was determined. Human TNBC cell lines were treated at increasing TMZ concentrations. mediated cytotoxicity was determined. Human TNBC cell lines were treated at increasing TMZ At 72 h post treatment, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay revealed concentrations. At 72 h post treatment, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) that cell viability decreased in a dose-dependent manner. HCC1937 cells displayed greater resistance assay revealed that cell viability decreased in a dose-dependent manner. HCC1937 cells displayed to TMZ than MDA-MB-231. A TMZ dose of 0.4 mM resulted in 20% cell viability in HCC1937 cells and greater resistance to TMZ than MDA-MB-231. A TMZ dose of 0.4 mM resulted in 20% cell viability 40% cell viability in MDA-MB-231 cells (Figure 1C). in HCC1937 cells and 40% cell viability in MDA-MB-231 cells (Figure 1C). Figure 1. Oncolytic adenovirus expressing mCherry (OAdmCherry) and temozolomide (TMZ) have Figure 1. Oncolytic adenovirus expressing mCherry (OAdmCherry) and temozolomide (TMZ) have a a cell-killing effect on human triple-negative breast cancer (TNBC) cells: (A) HCC1937 and MDA-MB- cell-killing effect on human triple-negative breast cancer (TNBC) cells: (A) HCC1937 and MDA-MB-231 231 cells were infected with OAdmCherry or adenovirus expressing green fluorescent protein cells were infected with OAdmCherry or adenovirus expressing green fluorescent protein (AdGFP) (AdGFP) at different multiplicity of infection concentrations for 72 h. Crystal violet staining was used at different multiplicity of infection concentrations for 72 h. Crystal violet staining was used to to evaluate cytopathic effect (CPE). A representative staining of three independent experiments is evaluate cytopathic effect (CPE). A representative staining of three independent experiments is shown. shown. (B) Relative cell survival was calculated by measuring the absorbance of solubilized dye at (B) Relative cell survival was calculated by measuring the absorbance of solubilized dye at 590 nm. 590 nm. (C) HCC1937 and MDA-MB-231 cells were treated with TMZ at different concentrations for (C) HCC1937 and MDA-MB-231 cells were treated with TMZ at different concentrations for 72 h. 72 h. Cell survival was calculated by MTT assay. Results represent the mean of three repeated Cell survival was calculated by MTT assay. Results represent the mean of three repeated measurements measurements ± standard deviation (SD; error bars) (* p < 0.05). ± standard deviation (SD; error bars) (* p < 0.05).PDF Image | Temozolomide Enhances Triple-Negative Breast Cancer Virotherapy
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