Journal: Scientific Reports
Article Title: Y-box binding protein 1/cyclin A1 axis specifically promotes cell cycle progression at G 2 /M phase in ovarian cancer
doi: 10.1038/s41598-024-72174-9
Figure Lengend Snippet: Effects of YBX1 siRNA on the expression levels of YBX1, cyclin A1, cyclin A2, cyclin B1, cyclin D1, cyclin E1, CDC6, and CDC20 mRNAs in ovarian cancer cell lines. ( A ) The eight selected genes are listed. Correlations of YBX1 with other genes are presented according to Spearman scores based on gene ontology analysis (TCGA database, n = 316) of HGSC, and the chromosomal locations of the 8 genes are indicated. The right panel of the illustrated model shows the sites of action of 7 cell cycle-related genes in the cell cycle. ( B ) RT-qPCR analysis showing mRNA expression levels of cell cycle-related genes in ovarian cancer cells. Student’s t-tests were used to compare differences between YBX1 siRNA#1-transfection groups and the control group. * p < 0.05; ** p < 0.01. Error bars indicate standard deviations for 3 independent experiments. ( C ) RT-qPCR analysis showing mRNA expression levels of 7 genes in ovarian cancer cell lines following transfection with YBX1 siRNA#1 for 3 days. Expression levels were expressed as ratios relative to the expression in the corresponding negative control based on 3 independent assays. Student’s t-tests were used to compare differences between YBX1 siRNA#1-transfection groups and the control group. * p < 0.05; ** p < 0.01. Error bars indicate standard deviations for 3 independent experiments. A heatmap plot of cell cycle-related genes from the comparison of the YBX1 siRNA#1-transfection group and the control group was presented. High expression levels in the YBX1 siRNA#1-transfection group are indicated in red, and low expression levels are indicated in blue. ( D ) JHOC-5 cells were transfected with luciferase reporter constructs containing the 1166 bp 5′-flanking region of the cyclin A1 gene with YBX1 siRNA#1 for 2 days. The firefly luciferase activity was normalized to the Renilla luciferase activity. Student’s t-tests were used to compare differences between YBX1 siRNA#1-transfection groups and the control group. ** p < 0.01. Error bars indicate standard deviations for 3 independent experiments. (E) Schematic representation of potential YBX1 binding sites (Black boxes) and primer locations (arrows) used for ChIP assay in the promoter region of Cyclin A1 gene. (F) The YBX1 binding to the cyclin A1 promoter by ChIP assay in JHOC-5 cells.
Article Snippet: Antibodies against CDC20 (cat. no. NB100-59828) were purchased from Novus Biologicals (Centennial, CO, USA).
Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Negative Control, Comparison, Luciferase, Construct, Activity Assay, Binding Assay
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