Fig. 5

Loss of Oct4A reduced migratory and proliferative abilities in HEY cells while increasing chemosensitivity to cisplatin treatment. a The migratory ability of Oct4A KD cells was determined by 24 h wound healing assay. Cells were grown as confluent monolayer prior to the insertion of a single wound and allowed to migrate for 24 h. Images of the wounds were taken at t = 0 h and t = 24 h post wound insertion and are representative of three independent experiments. Images are at 100x magnification and scale bar represents 50 μM. b 24 h migration expressed as a percentage of wound closure compared to t = 0 h using an ocular micrometer. Data is presented as the mean ± SEM of three independent experiments. Significance is indicated by **p < 0.01 as determined by One-Way ANOVA compared to vector control. c qPCR analysis of MMP2 mRNA expression in Oct4A KD monolayer cells. Relative quantification of MMP2 mRNA expression in Oct4A knockdown cells was standardized to 18S housekeeping gene and normalized the vector control. d Proliferative potential of Oct4A KD cells was determined by MTT assay over a 72 h period. Proliferation rates are expressed as the percentage of cell growth compared to 24 h. Data is presented as the mean ± SEM of three individual experiments performed with 6 replicates. Significance at t = 72 h is indicated by *p < 0.05 and ***p < 0.001 as determined by Two-way ANOVA compared to HEY Vector Control. e mRNA expression of Oct4A in ovarian cancer cell lines following cisplatin treatment. Relative quantification of Oct4A mRNA expression in ovarian cancer cells was standardized to 18S housekeeping gene and normalized to untreated controls for each cell line. Data is presented as the mean ± SEM of three independent experiments. Significance is indicated by ***p < 0.001, **p < 0.01 and *p < 0.05 compared to the respected untreated control for each cell line as determined by student's test. f Loss of Oct4A increased HEY cell sensitivity to 3 day cisplatin treatment as determined by MTT assay. Data is presented as the mean ± SEM of three independent experiments performed in triplicate and expressed as a percentage of cell survival compared to untreated cells for each group. Significance is indicated by *p < 0.05 and ***p < 0.001 for both Oct4A KD1 and Oct4A KD2 at 1 μg/ml and 2 μg/ml as determined by Two-way ANOVA compared to vector control cells