Journal: Journal of Advanced Research

Article Title: Genomic landscape of paired primary and peritoneal metastatic lesions in gastric cancer highlights evolutionary dynamics and mutational drivers

doi: 10.1016/j.jare.2025.05.043

Figure Lengend Snippet: Roles of DEK in gastric cancer stemness and peritoneal metastasis. (A) Correlation analysis between DEK expression and cancer stem cell (CSC) signature in the TCGA gastric cancer cohort. (B) Detailed correlations between DEK and the 16 individual members of the CSC signature. (C) Correlations between DEK and two key CSC markers, SOX9 (right) and YAP1 (left), in the TCGA cohort. (D) Assessment of DEK levels in AGS cells transfected with DEK-encoding lentivirus (DEK-OE) or vector, along with their expression levels of YAP1 and SOX9 . (E) mRNA levels of DEK , YAP1 , and SOX9 measured in DEK-OE and control cells by RT-PCR, standardized to the corresponding expression levels in control cells. (F) Tumor sphere formation abilities in DEK-OE and control cells. Photographs of tumor sphere formation at 3-day intervals are shown on the left and quantitative analysis are on the right. (G) Schematic representation of the peritoneal metastasis model using DEK-OE or control tumor cells in mice. (H) Measurements of abdominal circumference (left) and body weight (right) changes in mice with DEK-OE versus control cells. (I-K) Visual (left) and quantitative assessment (right) of peritoneal metastasis models, showing general appearance (I), volume of ascites (J), and peritoneal implants (K) on day 20 for DEK-OE vs. control subcutaneous gastric cancer groups. (L) Kaplan-Meier survival curves for mouse models with DEK-OE versus control cells. (M−N) Protein (M) and mRNA (N) levels of DEK, YAP1, and SOX9 in peritoneal tumors from the mouse models. (O) Schematic representation of competition assays in the peritoneal model with DEK-OE and control cells. (P) Proportions of DEK-OE and control cells in the peritoneum of mice in the competition model, measured by flow cytometry.

Article Snippet: Paraffin-embedded sections were treated with goat serum to block non-specific binding and subsequently stained with anti-DEK antibody (Proteintech, Wuhan, China) and anti-YAP1 antibody (Proteintech, Wuhan, China) using an IHC staining kit (Zsbio, Beijing, China).

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry

Journal: Neoplasia (New York, N.Y.)

Article Title: The DEK Oncoprotein Functions in Ovarian Cancer Growth and Survival 1

doi: 10.1016/j.neo.2018.10.005

Figure Lengend Snippet: Decreasing DEK levels sensitizes ovarian cancer cell lines to chemotherapy. (Ai) OVCAR8 or CAOV3 ovarian cancer cells were transfected with siRNA targeting control sequence or siDEK for 24 hours. Immunoblotting was performed on cell lysates for DEK expression. (Aii) After OVCAR8 and CAOV3 cells were transfected with siControl or siDEK for 24 hours, cells were treated for an additional 48 hours with cisplatin or panobinostat and assessed for cell viability by MTT assay. Statistical analysis was performed by ANOVA with Tukey's post hoc test; * P < .05, ** P < .01, *** P < .001, **** P < .0001 versus siControl. (Bi) CAOV3, OVCAR8, and OVCAR3 cells were infected with a lentivirus expressing a short hairpin RNA targeting a control sequence (shControl1) or DEK nt 1165-1185 (shDEK1). Immunoblotting was performed on cell lysates 72 hours after infection for DEK expression. (Bii) Twenty-four hours after infection with shControl1 or shDEK1, cells were treated with cisplatin, doxorubicin, or panobinostat for an additional 48 hours and assessed for cell viability using an MTT assay. Statistical analysis was performed by ANOVA with Tukey's post hoc test; * P < .05, ** P < .01, *** P < .001, **** P < .0001 versus shControl1. (Ci) Immunoblotting was performed on cell lysates following infection with lentiviral constructs with shControl2, shDEK2 (nt 860-880), or shDEK3 (nt 1192-1216). (Cii) Twenty-four-hour infection with shControl2, shDEK2, or shDEK3 followed by treatment with indicated therapies. Statistical analysis was performed by ANOVA with Tukey's post hoc test; * P < .05, ** P < .01, *** P < .001, **** P < .0001 versus shControl2. (Di) Following lentiviral infection with shControl1 or shDEK1, stable polyclonal cell lines were generated after selection with 0.25 μg/ml of puromycin. Growth curves were generated by counting total number of cells every 24 hours for 6 days. Statistical analysis was performed by ANOVA with Bonferroni post hoc test. ** P < .01, *** P < .001, **** P < .0001 versus shControl1. (Dii) Growth curves were generated in stable lines expressing shControl2, shDEK2, or shDEK. Statistical analysis was performed by ANOVA with Bonferroni post-hoc test. ** P < .01,*** P < .001, **** P < .0001 versus shControl2.

Article Snippet: For transient transfections, CAOV3 or OVCAR8 cells were grown to approximately 50% confluence and transfected using Lipofectamine 2000 (ThermoFisher) transfection reagent and 30 pmol of siRNA targeting DEK (Santa Cruz Biotechnology) or control siRNA (Santa Cruz Biotechnology).

Techniques: Transfection, Control, Sequencing, Western Blot, Expressing, MTT Assay, Infection, shRNA, Construct, Generated, Selection