Journal: American Journal of Translational Research

Article Title: Reduction of COX-2 through modulating miR-124/SPHK1 axis contributes to the antimetastatic effect of alpinumisoflavone in melanoma

doi:

Figure Lengend Snippet: AIF represses COX-2 expression by modulating miR-124/SPHK1 signaling. Cells were pretreated for 24 hours with AIF (10 μM unless otherwise noted). A. AIF treatment dose-dependently decreased SPHK1 protein levels in melanoma cells. B. AIF treatment dose-dependently increased mIR-124 expression in melanoma cells. C. MiR-124 knockdown significantly attenuated the suppressive effects of AIF on SPHK1 expression, as demonstrated by western blot analysis. D. Overexpression of miR-124 significantly decreased SPHK1 transcription, as assessed by the luciferase reporter assay. E. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated the suppressive effect of AIF on COX-2 mRNA expression. F. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated the suppressive effect of AIF on COX-2 protein expression. **P < 0.01 vs. control, ^^P < 0.01 vs. AIF.

Article Snippet: Silencing COX-2 or SPHK1 in melanoma cells The siRNA oligos for COX-2 or SPHK1 gene knockdown were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Expressing, Knockdown, Western Blot, Over Expression, Luciferase, Reporter Assay, Control

Journal: American Journal of Translational Research

Article Title: Reduction of COX-2 through modulating miR-124/SPHK1 axis contributes to the antimetastatic effect of alpinumisoflavone in melanoma

doi:

Figure Lengend Snippet: MiR-124/SPHK1 signaling is involved in the antimetastatic effect of AIF. Cells were pretreated for 24 hours with AIF (10 μM if not otherwise noted). A. COX-2 overexpression attenuated the effect of AIF on melanin content in melanoma cells. B. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated the effect of AIF on PpIX accumulation. C. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated the effect of AIF on TG2 activity. D. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated AIF-mediated inhibition of cell migration. E. Both MiR-124 knockdown and SPHK1 overexpression significantly attenuated AIF-mediated inhibition of cell invasion. **P < 0.01.

Article Snippet: Silencing COX-2 or SPHK1 in melanoma cells The siRNA oligos for COX-2 or SPHK1 gene knockdown were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Over Expression, Knockdown, Activity Assay, Inhibition, Migration

Journal: American Journal of Translational Research

Article Title: Reduction of COX-2 through modulating miR-124/SPHK1 axis contributes to the antimetastatic effect of alpinumisoflavone in melanoma

doi:

Figure Lengend Snippet: AIF suppresses B16-F10 murine melanoma cell lung metastasis in vivo. Xenograft-bearing mice were treated for 24 days with vehicle or AIF (20 or 50 mg/kg/day) by intragastric administration. Representative images of lung metastasis and the number of metastatic nodules in the lungs are shown. A. AIF significantly suppressed lung metastasis in vivo. B. AIF increased miR-124 expression in vivo. C. AIF treatment decreased expression levels of COX-2 and SPHK1 in vivo. *P < 0.05 (n = 8).

Article Snippet: Silencing COX-2 or SPHK1 in melanoma cells The siRNA oligos for COX-2 or SPHK1 gene knockdown were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: In Vivo, Expressing

Journal: Molecular Oncology

Article Title: Co‐administration phenoxodiol with doxorubicin synergistically inhibit the activity of sphingosine kinase‐1 (SphK1), a potential oncogene of osteosarcoma, to suppress osteosarcoma cell growth both in vivo and in vitro

doi: 10.1016/j.molonc.2012.04.002

Figure Lengend Snippet: Inhibition of SphK1 sensitizes doxorubicin induced U2OS cell growth suppression. U2OS cells were either left untreated (Ctrl) or exposed to phenoxodiol (Pxd, 5 μg/ml), doxorubicin (Dox, 0.25 μM) or both (Pxd + Dox) ...

Article Snippet: 10 μl/ml of lentiviral particles containing SphK1 shRNA (sc‐44114‐V, Santa Cruz Biotech, Santa Cruz, CA) were added to the cells for 48 h, cell culture medium then is replaced by fresh medium and cells were cultured for another 24 h, stable clones expressing SphK1 shRNA were selected by puromycin (1 μg/ml).

Techniques: Inhibition

Journal: Molecular Oncology

Article Title: Co‐administration phenoxodiol with doxorubicin synergistically inhibit the activity of sphingosine kinase‐1 (SphK1), a potential oncogene of osteosarcoma, to suppress osteosarcoma cell growth both in vivo and in vitro

doi: 10.1016/j.molonc.2012.04.002

Figure Lengend Snippet: Identification SphK1 as a potential oncogene in OS. Western blots results shows the expression level of SphK1 in fresh removed five clinical OS tissues from and 2 normal osteoblast tissues (N) (A). Immunohistochemistry results confirmed the up‐regulation ...

Article Snippet: 10 μl/ml of lentiviral particles containing SphK1 shRNA (sc‐44114‐V, Santa Cruz Biotech, Santa Cruz, CA) were added to the cells for 48 h, cell culture medium then is replaced by fresh medium and cells were cultured for another 24 h, stable clones expressing SphK1 shRNA were selected by puromycin (1 μg/ml).

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: Molecular Oncology

Article Title: Co‐administration phenoxodiol with doxorubicin synergistically inhibit the activity of sphingosine kinase‐1 (SphK1), a potential oncogene of osteosarcoma, to suppress osteosarcoma cell growth both in vivo and in vitro

doi: 10.1016/j.molonc.2012.04.002

Figure Lengend Snippet: Proposed signaling pathways involved in this study. SphK1 is a potential key oncogene for OS. Phenoxodiol and doxorubicin synergistically inhibited SphK1 activity to increase ceramide production. Increased ceramide then induces PP1/Akt association to ...

Article Snippet: 10 μl/ml of lentiviral particles containing SphK1 shRNA (sc‐44114‐V, Santa Cruz Biotech, Santa Cruz, CA) were added to the cells for 48 h, cell culture medium then is replaced by fresh medium and cells were cultured for another 24 h, stable clones expressing SphK1 shRNA were selected by puromycin (1 μg/ml).

Techniques: Activity Assay