Mitotane cytotoxicity and metabolism after CYP11B1 modulation in H295R cell line.

<p>(A) Mitotane cytotoxicity was not influenced by CYP11B1 interference (p = ns, comparing IC50 doses of CYP11B1 siRna cells and non-targeting siRna control cells). Three replicate wells for each experiment (n = 2) were used to determine each data point. (B) CYP11B1 modulation did not influence uptake of mitotane and its metabolization (p = ns, comparing drug levels of CYP11B1 siRna cells and non-targeting siRna control cells). Four independent experiments were used to determinate each data point, measured in HPLC-UV.</p

Mitotane metabolites in ACC cells: Uptake, bioavailability and metabolization.

<p>Intracellular and extracellular concentration of mitotane and its metabolites in H295R, measured by means of HPLC–UV after metyrapone treatment. Six independent experiments were used to determinate each data point.</p

Effects of mitotane and 20 μM metyrapone on H295R cell viability.

<p>Three technical replicate wells for each experiment (n = 3) were used to determine each data point. P = ns comparing mitotane treatment vs. combined treatment with mitotane and metyrapone.</p

CYP11B1 expression after treatment with mitotane and metyrapone, or both.

<p>Data obtained from two independent experiments with two replicates for each experiment, and expressed as fold changes compared to β-actin expression (2-ΔΔCt). A > 2 fold increase was considered significant.</p

Efficiency of siRna transfection and silencing.

<p>(A) H295R cells transfected with a fluorescent siRna indicator. (B) CYP11B1 gene expression in CYP11B1 siRna cells and non-targeting siRna cells. Data obtained from two independent experiments with two replicates for each experiment. (C) Cortisol levels in CYP11B1 siRna cells and non-targeting siRna control cells. Four independent experiments were used to determine each data point.</p

Effects of 25 μM mitotane and 20 μM metyrapone on cortisol levels.

<p>Four independent experiments were used to determine each data point. Metyrapone or mitotane treatment vs. untreated cells, P <0.05. Combined treatment with mitotane and metyrapone vs. untreated cells. P = 0.005.</p

The major issue of oncological treatment administration at the end of life: a retrospective study - supplementary material

Supplementary table S1: Clinicopathological features of study case seriesSupplementary Table S3: Univariate analysis of variable potential predicting end-of-life therapy</p

Kaplan-Meier estimates of overall survival according to a combined risk factors model with Argiris factors and CTCs.

<p>Continuous line indicates absence of both risk factors; small dotted line indicates the presence of only one of the two risk factors; large dotted line indicates the presence of both risk factors.</p

Example of CTCs analysis in a patient with mediastinal and axillary nodal metastases from an oropharyngeal squamous cell carcinoma.

<p>(A) the CellSearch output of baseline CTC analysis showing two CTCs with heterogeneous EGFR expression. (B) Timeline of CTC analysis and treatments. (C) Correlative imaging analysis by CT/PET at baseline and after chemotherapy. In this patient 3 CTCs were detected at baseline. After 4 cycles of a chemotherapy, CTC number rised to 9 suggesting progressive disease then confirmed by CT/PET imaging.</p

Association between the presence of CTCs before starting a new line of chemotherapy and response to treatment.

<p>Higher response rate is observed in CTC-negative patients at baseline (A). Dynamic variation of CTCs numbers before and after treatment in patients (n = 10) with at least two determinations and at least one CTC at any time point. CTCs changes did not correlate with tumor response (B).</p