human serous ovarian cancer skov3 cell line  (iCell Bioscience Inc)

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: The effect of ectopic expression of p53 mutants on cell migration and invasion in p53-null SKOV3 cells. ( A ) A western blot assay was performed to measure the p53 protein levels in stably transfected SKOV3 EV , SKOV3 R175 , SKOV3 R248 , and SKOV3 R273 cells. β-Actin was used as an internal control. The cells were seeded in uncoated chambers for the migration assay and incubated for 24 h or seeded in Matrigel-coated chambers for the invasion assay and incubated for 48 h. ( B ) A western blot assay was performed to measure the p53 protein levels after SKOV3 cells were transiently transfected with empty vector, wild-type p53, p53-R175, p53-R248, and p53-R273. β-Actin was used as an internal control. The SKOV3 cells were seeded in uncoated chambers for the migration assay and incubated for 24 h or seeded in Matrigel-coated chambers for the invasion assay and incubated for 48 h. Representative images show the migrating and invading cells. Results are the combined data (mean ± S.D.) from three independent experiments. *P < 0.05 compared with the SKOV3 EV group or the empty vector group.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Expressing, Migration, Western Blot, Stable Transfection, Transfection, Control, Incubation, Invasion Assay, Plasmid Preparation

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: The effect of mutant p53 knockdown on mutant p53-induced ovarian cancer cell migration and invasion. ( A ) Stably transfected SKOV3 EV , SKOV3 R175 , SKOV3 R248 , and SKOV3 R273 cells were transfected with control or p53 siRNA, and then p53 protein levels were measured by western blot assay. β-Actin was used as an internal control. ( B ) After transfection with the siRNAs, the cells were seeded in uncoated chambers for the migration assay and incubated for 24 h. ( C ) After transfection with the siRNAs, the cells were seeded in Matrigel-coated chambers for the invasion assay and incubated for 48 h. Representative images show the migrating ( B ) or invading ( C ) cells. Results are the combined data (mean ± S.D.) from three independent experiments. *P < 0.05 compared with the control siRNA-transfected SKOV3 EV group and # P < 0.05 compared with control siRNA-transfected SKOV3 R175 , SKOV3 R248 , and SKOV3 R273 group, respectively.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Mutagenesis, Knockdown, Migration, Stable Transfection, Transfection, Control, Western Blot, Incubation, Invasion Assay

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: The effect of mutant p53 on the expression of migration/invasion-associated genes in ovarian cancer cells. ( A ) Real-time RT-PCR shows the mRNA levels of four selected genes (S1PR1, EDN2, THBS1, and HB-EGF) in stably transfected SKOV3 EV and SKOV3 R248 cells. *P < 0.05 compared with the SKOV3 EV group. ( B ) SKOV3 cells were transiently transfected with empty vector, wild-type p53, and p53-R248. Real-time RT-PCR was performed to evaluate the mRNA levels of four selected genes (S1PR1, EDN2, THBS1, and HB-EGF) in the transiently transfected SKOV3 cells. *P < 0.05 compared with the empty vector group. ( C ) After transfection with p53 siRNA or control siRNA, real-time RT-PCR was performed to measure the mRNA levels of S1PR1, EDN2, THBS1, and HB-EGF in stably transfected SKOV3 EV and SKOV3 R248 cells. *P < 0.05 compared with the control siRNA-transfected SKOV3 EV group and # P < 0.05 compared with control siRNA-transfected SKOV3 R248 group. All expression levels were normalized to GAPDH. Results are the combined data (mean ± S.D.) from three independent experiments.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Mutagenesis, Expressing, Migration, Quantitative RT-PCR, Stable Transfection, Transfection, Plasmid Preparation, Control

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: Involvement of Rad21 in mutant p53-induced ovarian cancer cell invasion. ( A ) The mRNA levels of S1PR1, EDN2, THBS1, and HB-EGF were measured by real-time RT-PCR after transfection with control siRNA or Rad21 siRNA in SKOV3 EV and SKOV3 R248 cells. All expression levels were normalized to GAPDH. ( B ) The protein levels of S1PR1 and THBS1 were determined by western blot analysis after transfection with Rad21 siRNA (left panel) and p53 siRNA (right panel) in SKOV3 EV and SKOV3 R248 cells. β-Actin was used as an internal control. ( C ) After transfection with siRNA, the cells were seeded in uncoated chambers for the migration assay and incubated for 24 h. ( D ) After transfection with siRNA, the cells were in Matrigel-coated chambers for the invasion assay and incubated for 48 h. Representative images show the migrating ( C ) and invading ( D ) cells. Results are the combined data (mean ± S.D.) from three independent experiments. *P < 0.05 compared with the control siRNA-transfected SKOV3 EV group and # P < 0.05 compared with control siRNA-transfected SKOV3 R248 group.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Mutagenesis, Quantitative RT-PCR, Transfection, Control, Expressing, Western Blot, Migration, Incubation, Invasion Assay

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: The effect of the mutant p53 and Rad21 interaction on the transcriptional activation of S1PR1 and THBS1 in ovarian cancer cells. ( A ) The protein interaction of Rad21 with mutant p53-R248 was determined by an immunoprecipitation assay in stably transfected SKOV3 EV and SKOV3 R248 cells. The p53 protein was immunoprecipitated using an anti-p53 antibody. The reverse immunoprecipitation (IP) was also performed using an anti-Rad21 antibody. Proteins were analyzed by western blotting using anti-p53 and anti-Rad21-specific antibodies. Data are representative of three different experiments. ( B ) The putative Rad21-binding site in the S1PR1 and THBS1 genes. ( C ) After transient transfection with mutant p53-R248 or empty vector in SKOV3 cells, ChIP assays were performed. Cross-linked DNA fragments were immunoprecipitated with the anti-Rad21 antibody, and the purified DNA was amplified by PCR using gene-specific primers for the Rad21-binding sites of S1PR1 or THBS1. ( D ) After transfection with control siRNA or p53 siRNA in stably transfected SKOV3 EV and SKOV3 R248 cells, respectively, ChIP assays were performed. Cross-linked DNA fragments were immunoprecipitated with the anti-Rad21 antibody, and the purified DNA was amplified by PCR using gene-specific primers for the Rad21-binding sites of S1PR1 or THBS1. The binding site was not detected when normal IgG was used or the antibody was omitted from the immunoprecipitation (IP) step. Data are representative of three different experiments. *P < 0.05 compared with the empty vector group or the SKOV3 EV group and # P < 0.05 compared with control siRNA transfected-SKOV3 R248 group.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Mutagenesis, Activation Assay, Immunoprecipitation, Stable Transfection, Transfection, Western Blot, Binding Assay, Plasmid Preparation, Purification, Amplification, Control

Journal: Scientific Reports

Article Title: Mutant p53 stimulates cell invasion through an interaction with Rad21 in human ovarian cancer cells

doi: 10.1038/s41598-017-08880-4

Figure Lengend Snippet: Involvement of S1PR1 in mutant p53-induced ovarian cancer cell invasion. ( A ) After transfection with control siRNA or S1PR1 siRNA for 24 h, the cells were seeded in Matrigel-coated chambers for invasion assay and incubated for 48 h. ( B ) After transfection with control siRNA or THBS1 siRNA for 24 h, the cells were seeded in Matrigel-coated chambers for invasion assay and incubated for 48 h. Representative images show the invading cells. Results are the combined data (mean ± S.D.) from three independent experiments. *P < 0.05 compared with the control-siRNA transfected SKOV3 EV group and # P < 0.05 compared with control siRNA-transfected SKOV3 R248 group.

Article Snippet: The high grade serous human ovarian cancer cell line SKOV3 was originally from the American Type Culture Collection.

Techniques: Mutagenesis, Transfection, Control, Invasion Assay, Incubation