Journal: Cancer letters

Article Title: CD24 suppresses malignant phenotype by downregulation of SHH transcription through STAT1 inhibition in breast cancer cells.

doi: 10.1016/j.canlet.2015.12.013

Figure Lengend Snippet: Fig. 4. CD24 inhibits SHH transcription through suppression of STAT1. (A) Heatmaps of selected genes identified as related to the SHH and STAT family by DNA microarray analysis of MCF-7 or HCC1937 cells. (B) STAT1 mRNA expression in CD24 siRNA-transfected MCF-7 and HCC1937 cells was measured by real time RT-PCR. STAT1 and pSTAT1 expressions in CD24 siRNA-transfected MCF-7 and HCC1937 cells were estimated by western blotting. (C) STAT1 mRNA expression in CD24-expressing Hs578T cells was evaluated by real time RT-PCR. STAT1 and pSTAT1 expressions in CD24 forced-expressing Hs578T cells were estimated by western blotting. (D) SHH mRNA and protein ex- pressions in CD24 siRNA-transfected cells, and STAT1 siRNA and CD24 siRNA-co-transfected cells in MCF-7 and HCC1937 were analyzed by real time RT-PCR. (E) The total number of migrating cells in MCF-7 and HCC1937 cells transfected with control, CD24, or both CD24 and STAT1 siRNA was evaluated by matrigel invasion assay. (F) Anchorage- independent proliferation of CD24 siRNA-transfected cells, and STAT1 and CD24 siRNA-co-transfected cells at three weeks following plating in agar (G) Tumorigenicity of CD24 and STAT1 siRNA-co-transfected cells compared with tumorigenicity of CD24 siRNA-transfected cells. Approximately 5.0 × 105 MCF-7 cells and HCC1937 cells were injected subcutaneously into each side of the dorsal region of NOD/SCID mice (n = 8 per group). *p < 0.05, **p < 0.01. Bar represents SD.

Article Snippet: Empty vector control (eGFP-Control) and STAT1 plasmids (eGFPSTAT1) were purchased from Addgene (Cambridge, MA, USA).

Techniques: Microarray, Expressing, Transfection, Quantitative RT-PCR, Western Blot, Control, Invasion Assay, Injection

Journal: Cancer letters

Article Title: CD24 suppresses malignant phenotype by downregulation of SHH transcription through STAT1 inhibition in breast cancer cells.

doi: 10.1016/j.canlet.2015.12.013

Figure Lengend Snippet: Fig. 5. STAT1 over-expression induces upregulation of SHH transcription. (A) Expression of STAT1 mRNA and protein in MCF-7 and HCC1937 cells transfected with STAT1- expressing plasmid was analyzed by real time RT-PCR and western blotting. (B) SHH mRNA and protein expressions in MCF-7 and HCC1937 cells transfected with STAT1 plasmid were measured by real time RT-PCR and western blotting. (C) Invasiveness of MCF-7 and HCC1937 cells transfected with STAT1 plasmid was determined by matrigel invasion assay. (D) MMP2 and MMP9 mRNA and protein in STAT1 over-expressing cells were analyzed by real time RT-PCR and western blotting. (E) Anchorage- independent proliferation of STAT1 plasmid-transfected MCF-7 cells at three weeks following plating in agar. *p < 0.05, **p < 0.01. Bar represents SD. (F) Representative pictures of immunofluorescence (IF) staining of surgically resected breast cancer specimens for CD24 (green; Alexa488) and pSTAT1 (red; Alexa594) (F-a), or pSTAT1 (green; Alexa488) and SHH (red; Alexa594) (F-b). Original magnification is ×400. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Empty vector control (eGFP-Control) and STAT1 plasmids (eGFPSTAT1) were purchased from Addgene (Cambridge, MA, USA).

Techniques: Over Expression, Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Invasion Assay, Staining

Journal: Cancer letters

Article Title: CD24 suppresses malignant phenotype by downregulation of SHH transcription through STAT1 inhibition in breast cancer cells.

doi: 10.1016/j.canlet.2015.12.013

Figure Lengend Snippet: Fig. 6. Overall schematic of CD24/STAT1/SHH signaling in breast cancer. CD24 in- hibits STAT1 activation in breast cancer. In the absence of CD24 (i.e. CD24−cells), transcription of STAT1 is activated, and STAT1 then upregulates SHH transcription to activate Hedgehog signaling in a ligand-dependent manner. Hedgehog signaling triggers induction of malignant characteristics including anchorage-independent pro- liferation, invasion, and tumorigenicity.

Article Snippet: Empty vector control (eGFP-Control) and STAT1 plasmids (eGFPSTAT1) were purchased from Addgene (Cambridge, MA, USA).

Techniques: Activation Assay