cell culture human skov3 eoc cells  (ATCC)

Journal: Annals of Translational Medicine

Article Title: Inhibition of sphingosine kinase 2 down-regulates ERK/c-Myc pathway and reduces cell proliferation in human epithelial ovarian cancer

doi: 10.21037/atm-20-6742

Figure Lengend Snippet: SphK2 inhibition suppresses the growth of EOC cells in vitro and in vivo. (A) SKOV3 cells transfected with control shRNA or SphK2 shRNA. Expression of SphK2 mRNA levels determined by PCR and normalized to GAPDH mRNA. (B) Protein levels of SphK2 as determined by Western blot. (C) A significant reduction in cell growth rate in the SphK2 shRNA group is observed by the cellular proliferation assay in ovarian cancer cells. (D) Representative images of tumors in subcutaneous ovarian cancer xenograft model. Mean tumor volume (E) and weight (F) in the SphK2 shRNA group are significantly reduced compared with the control group (8 mice in each group). The experiments were repeated 3 times. *, P<0.05; **, P<0.01. SphK2, sphingosine kinase 2; EOC, epithelial ovarian cancer; shRNA, short hairpin RNA; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Cell lines and culture conditions Human EOC cell lines SKOV3 and OVCAR3 were obtained from American Type Culture Collection (ATCC).

Techniques: Inhibition, In Vitro, In Vivo, Transfection, Control, shRNA, Expressing, Western Blot, Proliferation Assay

Journal: Annals of Translational Medicine

Article Title: Inhibition of sphingosine kinase 2 down-regulates ERK/c-Myc pathway and reduces cell proliferation in human epithelial ovarian cancer

doi: 10.21037/atm-20-6742

Figure Lengend Snippet: SphK2 inhibition in EOC cells causes an arrest in the G0/G1 phase of the cell cycle. (A) Representative flow cytometry charts of SKOV3 cells transfected with SphK2 shRNA or Ctrl shRNA and stained by PI. The percentage of cells in each cell cycle population was quantified. (B) Western blot analysis of cyclin D1 and p-Rb in ovarian cancer cells transfected with SphK2 shRNA or Ctrl shRNA. Right panels show that densitometric analysis of cyclin D1 and p-Rb (normalized to GAPDH) is reduced in SKOV3. (C) Bar graphs showing the percentage of apoptotic cells of SKOV3 transfected with SphK2 shRNA or Ctrl shRNA. The experiments were repeated 3 times. *, P<0.05. SphK2, sphingosine kinase 2; EOC, epithelial ovarian cancer; Ctrl, control; shRNA, short hairpin RNA; PI, propidium iodide; p-Rb, phosphate retinoblastoma; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Cell lines and culture conditions Human EOC cell lines SKOV3 and OVCAR3 were obtained from American Type Culture Collection (ATCC).

Techniques: Inhibition, Flow Cytometry, Transfection, shRNA, Staining, Western Blot, Control

Journal: Annals of Translational Medicine

Article Title: Inhibition of sphingosine kinase 2 down-regulates ERK/c-Myc pathway and reduces cell proliferation in human epithelial ovarian cancer

doi: 10.21037/atm-20-6742

Figure Lengend Snippet: SphK2 suppression induces an expression profile correlating to MAPK signaling pathway and MYC targets. (A) Heatmap of 100 genes with most significant changes in gene expression of SKOV3 cells transfected with SphK2 shRNA or Ctrl shRNA. Blue and red colors represent low and high relative gene expression levels, respectively. (B) List of the ten most significant KEGG gene set pathways correlating to gene expression changes resulting from SphK2 knockdown in SKOV3 cells. (C,D,E,F) GSEA shows significant association between SphK2 level and several gene sets, including MAPK signaling pathway (C), ERK MAPK targets (D), MYC target V1 (E) and MYC target V2 (F). SphK2, sphingosine kinase 2; MAPK, mitogen-activated protein kinase; MYC, myelocytomatosis; shRNA, short hairpin RNA; KEGG, Kyoto Encyclopedia of Genes and Genomes; ERK, extracellular regulated protein kinases; GSEA, gene set enrichment analysis.

Article Snippet: Cell lines and culture conditions Human EOC cell lines SKOV3 and OVCAR3 were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Gene Expression, Transfection, shRNA, Knockdown