Fig. 5: CDK4/6 inhibition inhibits the growth of JEG3R cells in vitro and in vivo.

A JEG3 and JEG3R cells silenced (siCDK4/6) or not (NT) for CDK4 and six using siRNAs were subjected to clonogenic growth assay. Top panel: representative picture of clonogenic assay dishes stained with Crystal Violet. Bottom panel: colony numbers as a bar graph of mean ± SEM of three independent experiments performed in triplicates. B Normal placental (PLC), JEG3 and JEG3R cells were grown in the presence of increasing concentrations of Palbociclib and cell survival determined at 96 h using Crystal violet staining. Graph represents mean ± SEM of biological triplicates with n = 2 normalised to the corresponding untreated control. C JEG3R cells were treated with 23 µM MTX or DMSO (diluent control) together with increasing concentrations of Palbociclib. Cell survival was determined by Crystal Violet staining. Data are mean ± SEM of biological triplicates with n = 4 normalised to the corresponding Palbociclib untreated control. D, E JEG3R cells were injected at the uterine horn of nude mice. Mice received either Vehicle-only (VO) and Palbociclib (PALB-125mg/kg) or Methotrexate (MTX-1mg/kg). D Representative pictures from ex vivo uteri showing tumour burden. NT; animals not injected with tumour cells. E Blood collected on indicated days was analysed for β-hCG levels by ELISA. Data representative of experiments performed in duplicate. Statistics: Student t-test. ****p < 0.001, ***p < 0.005, **p < 0.01, *p < 0.05, ns non-significant.