mouse sod2 mrna  (TriLink)

Journal: Cell Death & Disease

Article Title: Targeting prooxidant MnSOD effect inhibits triple-negative breast cancer (TNBC) progression and M2 macrophage functions under the oncogenic stress

doi: 10.1038/s41419-021-04486-x

Figure Lengend Snippet: A , B The relapse-free survival (RFS) of breast cancer patients classified by low ( n = 2947) and high ( n = 1004) MCTS1 expression ( A ) or by low ( n = 1449) and high ( n = 2502) SOD2 expression ( B ) was evaluated using Kaplan–Meier (KM) Plotter analysis. C The correlation of MCTS1 and SOD2 gene expression in breast cancer patients ( n = 159) was analyzed using the Pawitan dataset in the Oncomine cancer profiling database. D–F SOD2 expression was analyzed in TissueScan cDNA arrays with normal breast tissue ( n = 11) and breast tumor biopsies ( n = 124) across distinctive tumor stages ( D , E ), molecular subtypes and TNM classifications ( F ). The SOD2 mRNA levels in tumor samples were normalized to internal ACTB (β-actin) mRNA levels and then compared with the levels in normal breast samples. Data are presented as the mean ± s.e.m. G KM Plotter was used to estimate the survival of patients with MCTS1 high / SOD2 high ( n = 37) expression compared with that of those with MCTS1 low / SOD2 low ( n = 60) expression using the Pawitan dataset in the Oncomine database. H , I RFS was estimated for ER-negative ( n = 434) ( H ) and basal-like ( n = 309) ( I ) breast cancer patients with MCTS1 high / SOD2 high or MCTS1 low / SOD2 low expression using KM Plotter. Statistical analysis was performed using the log-rank Mantel–Cox test ( A , B , and G–I ), Pearson product-moment correlation coefficient ( C ), Kruskal–Wallis test followed by Dunn’s multiple comparison test ( D ) and χ 2 test ( E , F ).

Article Snippet: Further characterization of SOD2 mRNA levels was conducted with breast cancer cDNA arrays (OriGene).

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: Targeting prooxidant MnSOD effect inhibits triple-negative breast cancer (TNBC) progression and M2 macrophage functions under the oncogenic stress

doi: 10.1038/s41419-021-04486-x

Figure Lengend Snippet: A MnSOD, Nrf2, and catalase levels were examined in normal MCF-10A cells and metastatic MDA-MB-231 cells. B The SOD2 mRNA levels in MCF-10A, MDA-MB-231, and HCC1395 cells without (control) or with MCT-1 overexpression were measured by qRT-PCR. C , D MnSOD, Nrf2 and catalase levels were assayed upon MCT-1 gene silencing (shMCT-1 #1, #2 or #3) in MCF-10A ( C ) and MDA-MB-231 ( D ) cells. E Upon cycloheximide (CHX) treatment for the indicated times, Nrf2 protein stability and its amount relative to that of internal ACTB (right) were compared in MDA-MB-231 cells (control vs. MCT-1). Representative images (left) from four independent experiments are shown. F , G NFE2L2 mRNA levels were analyzed across distinctive breast tumor stages ( F ) and molecular subtypes or TNM classifications ( G ) using TissueScan breast cancer cDNA arrays. The NFE2L2 mRNA levels in tumor samples were normalized to the internal ACTB mRNA level and then compared with the levels in normal breast samples. H-I KM Plotter analysis estimated relapse-free survival in basal-like ( H ) breast cancer patients with MCTS1 high / NFE2L2 high ( n = 155) expression versus those with MCTS1 low / NFE2L2 low ( n = 154) expression and in breast cancer patients with lymph node metastasis (LN positive) ( I ) stratified by a SOD2 high / NFE2L2 high ( n = 283) or SOD2 low / NFE2L2 low ( n = 283) signature. J The correlation of SOD2 and NFE2L2 expression in TNBC patients was analyzed using the Stickeler dataset ( n = 32) in the Oncomine database. Data ( B , E ) are presented as the mean ± s.e.m. Statistical analysis was performed using a two-tailed unpaired Student’s t -test ( B ), linear regression followed by an analysis of variance (ANOVA) ( E ), the χ 2 test ( F , G ), the log-rank Mantel–Cox test ( H , I ), and the Pearson product-moment correlation coefficient ( J ). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Further characterization of SOD2 mRNA levels was conducted with breast cancer cDNA arrays (OriGene).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Targeting prooxidant MnSOD effect inhibits triple-negative breast cancer (TNBC) progression and M2 macrophage functions under the oncogenic stress

doi: 10.1038/s41419-021-04486-x

Figure Lengend Snippet: A The amounts of Nrf2 and MnSOD in MDA-MB-231 cells (control vs. MCT-1) were examined upon IL-6 (50 ng/ml) stimulation for different intervals. B IL-6 knockdown (shIL-6) was confirmed in MDA-MB-231 cells (control vs. MCT-1). C The expression of M2 markers ( CD163 and MRC1 ) were examined after THP-1 macrophages were cocultured with MDA-MB-231 cells (control vs. MCT-1) with or without shIL-6. D Secreted IL-6 levels were measured to evaluate MDA-MB-231 cells with or without MnSOD knockdown (scramble vs. shMnSOD) and MCT-1 overexpression (control vs. MCT-1). E The correlation of SOD2 and IL6 expression in TNBC patients ( n = 151) was analyzed using the Gluck dataset in the Oncomine. F The correlation of SOD2 and IL6 expression in TNBC patients ( n = 29) was examined using TissueScan breast cancer cDNA arrays. G , H Nrf2, MnSOD, and IL-6 levels were assayed after exposure to DPI ( G ) or Rotenone (Rot) ( H ). I Mitochondrial ROS (mROS) levels were quantified in MDA-MB-231 cells (scramble vs. shMCT-1) with or without MnSOD overexpression (MOCK vs. MnSOD) using superoxide fluorescent probe MitoSOX. J mROS levels were quantified in MDA-MB-231 cells (control vs. MCT-1) with or without MnSOD silencing (scramble vs. shMnSOD #7 and #8). K , L The peroxidase ( K ) and MnSOD dismutase activities ( L ) of MDA-MB-231 cells with or without MCT-1 knockdown (scramble vs. shMCT-1) and MnSOD overexpression (MOCK vs. MnSOD). M H 2 O 2 release from MDA-MB-231 cells with or without MCT-1 knockdown (scramble vs. shMCT-1) and MnSOD overexpression (MOCK vs. MnSOD) was measured using Amplex Red. N Quantification of H 2 O 2 released by MDA-MB-231 cells (scramble vs. shMCT-1) in MOCK vs. MnSOD expression conditions upon IL-6 stimulation for 24 h. Data are presented as the mean ± s.e.m. Statistical analysis was performed using two-way analysis of variance (ANOVA) followed by Tukey–Kramer (C, D, I–K, M-N) or Newman–Keuls (L) post hoc analysis or the Pearson correlation coefficient ( E , F ). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Further characterization of SOD2 mRNA levels was conducted with breast cancer cDNA arrays (OriGene).

Techniques: Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: Targeting prooxidant MnSOD effect inhibits triple-negative breast cancer (TNBC) progression and M2 macrophage functions under the oncogenic stress

doi: 10.1038/s41419-021-04486-x

Figure Lengend Snippet: A MDA-MB-231 cells (1 × 10 6 ) with or without MCT-1 overexpression (control vs. MCT-1) and MnSOD knockdown (scramble vs. shMnSOD) were orthotopically implanted into the mammary fat pad of nude mice ( n = 6 ~ 9). Tumor growth rates were monitored weekly. B Representative tumor images (left) and tumor burdens (righ) were assessed at week 9 in xenograft mice. C Tumor mortality was analyzed after orthotopic injection of the indicated MDA-MB-231 cells into nude mice ( n = 8). D Immunohistochemistry was used to examine tumor-associated CD163(+) M2 macrophages (denoted by red arrowheads). Scale bar, 50 µm. E Flow cytometric analysis of M2 macrophages (CD206(+) in F4/80(+) populations) residing in tumors was performed. F The correlation of the SOD2 expression in invasive breast carcinoma patients of basal-like subtypes ( n = 191) in The Cancer Genome Atlas (TCGA) cohort with the infiltration of M2 macrophages was evaluated using TIMER 2.0 and analyzed by the CIBERSORT-ABS tool. Expression levels are indicated as log2 transcripts per million (TPM). G Schematic figure depicting that amplification of the IL-6/MCT-1/Nrf2/MnSOD signaling loop aggravates TNBC progression/stemness and enhances M2 functions. MnSOD silencing thereby abolishes TNBC growth and improves M1 macrophage-mediated phagocytosis. Data are presented as the mean ± s.e.m. Statistical analysis was performed using a two-tailed unpaired Student’s t -test ( A , D ), the log-rank Mantel–Cox test ( C ), two-way analysis of variance (ANOVA) followed by Tukey–Kramer post hoc analysis ( B , E ), and the Spearman’s rank correlation coefficient (Rho) ( F ). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Further characterization of SOD2 mRNA levels was conducted with breast cancer cDNA arrays (OriGene).

Techniques: Over Expression, Injection, Immunohistochemistry, Expressing, Amplification, Two Tailed Test

Journal: The American Journal of Pathology

Article Title: Hepatitis C and Alcohol Exacerbate Liver Injury by Suppression of FOXO3

doi: 10.1016/j.ajpath.2013.08.013

Figure Lengend Snippet: Effects of HCV and alcohol on FOXO3 transcriptional activity. A: Real-time RT-PCR for FOXO3-dependent genes, FOXO3, SOD2, Bim, and GADD45, and a control, non-FOXO target gene, SOD1, in RNA isolated from control (black bars) and 5-day HCV-infected (gray bars) Huh7.5 cells. n = 6 to 18 individual RNA preparations. P values by Wilcoxon signed-rank test for the comparison of infected and noninfected conditions were as follows: FOXO3, P < 0.001; SOD2, P < 0.001; BIM, P < 0.005; GADD45, P < 0.05; SOD1, P = 0.86. B: Time course of mRNA expression changes determined by real-time PCR for FOXO3 and its target genes, Bim and SOD2, in control (black bars) and HCV-infected (gray bars) cells treated with 50 mmol/L ethanol for the indicated times. N = 3 to 6 individual preparations. C: HCV-infected and noninfected Huh7.5 cells were incubated with 50 mmol/L ethanol for 48 hours. Western blots for FOXO3, SOD2, Bim, and β-actin (loading control) are presented for whole cell lysates prepared at different times of alcohol exposure. Similar results were obtained in four independent experiments. D: SOD2 protein levels were measured by densitometry analysis of immunoblots performed as in A. n = 4. E: Kinetics of SOD2 degradation after cycloheximide treatment, determined as in D (n = 3 experiments). F: pMiR-Target-Luciferase-SOD2 3′-UTR reporter expression in HCV-infected and noninfected Huh7.5 cells incubated where indicated with 50 mmol/L ethanol for 48 hours. n = 3. ∗P < 0.05, ∗∗P < 0.01.

Article Snippet: F shows the results obtained using the pMiR-Target-Luciferase-SOD2 3′-UTR reporter system (OriGene) to assess the presence of miRNAs able to suppress SOD2 mRNA translation.

Techniques: Activity Assay, Quantitative RT-PCR, Isolation, Infection, Expressing, Real-time Polymerase Chain Reaction, Incubation, Western Blot, Luciferase

Journal: The American Journal of Pathology

Article Title: Hepatitis C and Alcohol Exacerbate Liver Injury by Suppression of FOXO3

doi: 10.1016/j.ajpath.2013.08.013

Figure Lengend Snippet: Effects of alcohol feeding on mouse liver. Mice from the four genotypes were fed either a Lieber-DeCarli liquid alcohol diet or a control, alcohol-free liquid diet for 3 weeks, as described in Materials and Methods. A: Serum ALT values expressed for each mouse were expressed as fold elevation compared with WT mouse on the control diet. ALT in the ethanol-fed HCV/Sod2+/− group was significantly different from the other groups by analysis of variance (P < 0.001). White squares, pair fed; red circles, alcohol fed. B: Representative H&E-stained liver histopathological characteristics for the different groups. C: Western blot analysis for inflammation and apoptosis markers in 3-week alcohol-fed and control-fed mice. D: Higher magnification of features of the HCV/Sod2+/− alcohol-treated livers demonstrating ballooning degeneration (top panel) and lobular inflammation (bottom panel). Features are indicated by arrows. Casp, caspase.

Article Snippet: F shows the results obtained using the pMiR-Target-Luciferase-SOD2 3′-UTR reporter system (OriGene) to assess the presence of miRNAs able to suppress SOD2 mRNA translation.

Techniques: Staining, Western Blot

Journal: The American Journal of Pathology

Article Title: Hepatitis C and Alcohol Exacerbate Liver Injury by Suppression of FOXO3

doi: 10.1016/j.ajpath.2013.08.013

Figure Lengend Snippet: SOD2 and FOXO3 expression and distribution in alcohol-fed mouse livers. A: Correlation of ALT with postalcohol SOD2 level in liver. SOD2 expression relative to β-actin was determined by densitometry of Western blots and plotted versus serum ALT. Blue diamonds, control-fed; red circles, alcohol fed. B: Comparison of the effects of alcohol feeding on SOD2 protein abundance in liver for the four genotypes. Liver homogenates were analyzed by using Western blot analysis after 3 weeks of alcohol or control diet feeding. Each lane represents homogenate from a separate mouse. C: FOXO3 immunohistological characteristics are presented. For each genotype, paraffin sections of liver were stained for FOXO3; and for each diet, images are shown at low power (10× objective, left panels) and at higher magnification (right panels), as indicated. The arrow indicates a cell with cytosolic immunoreactivity for FOXO3. D: Quantitative analysis of proportion of hepatocytes displaying cytosolic FOXO3 staining. Black bars, control diet; gray bars, ethanol diet. N = 3 different mouse livers for each condition. ∗∗P < 0.01 compared with Sod2+/−.

Article Snippet: F shows the results obtained using the pMiR-Target-Luciferase-SOD2 3′-UTR reporter system (OriGene) to assess the presence of miRNAs able to suppress SOD2 mRNA translation.

Techniques: Expressing, Western Blot, Staining