Skip to main content
Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Chemoresistance is mediated by ovarian cancer leader cells in vitro

Fig. 6

Leader cells proliferate at the same rate in chemotherapy and do not arise due to cellular division and DNA replication, leader cell deficient lines are more susceptible to cisplatin treatment. A Flow cytometric analysis of Ki67 surface expression in LC+/− populations in ovarian cancer cell lines in the presence of cisplatin. B Cells were seeded at 300,000 cells/well in a 6-well plate, incubated for 18 h, and treated with 13 μg/ml cisplatin +/− aphidicolin (0.5 μg/ml) or tunicamycin (6 μg/ml) for 24 h. Following treatment, cells were collected and stained for Ki67 using a Ki67-BV786 antibody. GFP (indicative of KRT14 expression) and BV786 fluorescence was acquired using the BD LSRFortessa™ X-20 with data analysed using the FlowJo software (v10.5.0). C Analysis was performed using a non-parametric Mann-Whitney U test to determine statistical significance between groups; n = 2–4/cell line; ns = not significant. D Wild-type, LC-deficient (KRT14KO) and LC-enriched (KRT14OE) cells were seeded at 0.2 × 104 cells/0.1 ml/well in 96-well plates. Cells were synchronized in G0 by overnight incubation in serum-free media prior to treatment and treated with cisplatin concentrations ranging from 1.2–30 μg/ml for 48 h followed by the addition of cell viability dye alamarBlue™ (Invitrogen). Half maximal inhibitory (IC50) concentrations of cisplatin were calculated in GraphPad Prism (v9.0) and analysed by non-linear regression fit. E Long-term assessment of cellular proliferation in response to high doses of Cisplatin where 0.2 × 104 cells/0.1 ml/well wild-type, LC deficient (KRT14KO) and LC enriched (KRT14OE) cells were seeded onto RTCA xCELLigence E-plates and treated with 30 μg/ml cisplatin, cellular proliferation was monitored over 240-h and impedance readings taken every 15 min

Back to article page