Fig. 5: Effects of NanoFEN on cell cycle and stem-like features in vitro and in vivo on lung cancer spheroid cells (LCSCs) and CRC spheroid cells (CSCs). | Cell Death & Disease

Fig. 5: Effects of NanoFEN on cell cycle and stem-like features in vitro and in vivo on lung cancer spheroid cells (LCSCs) and CRC spheroid cells (CSCs).

From: A new bioavailable fenretinide formulation with antiproliferative, antimetabolic, and cytotoxic effects on solid tumors

Fig. 5

a Schematic representation of cell cycle regulators activated (black) or repressed (grey) arised from RPPA analysis of LCSCs treated with NanoFEN as described in Fig. 4a. b Immunoblot analysis of cell cycle regulators phospho-Aurora A-B-C, phospho-PLK1, p38, phospho-p38 and p16 on LCSC6 (left panel) and CSC7 (right panel) untreated and treated with NanoFEN at the IC50 concentration (4.9 μM and 0.2 μM respectively) for 48 h. c Flow cytometry analysis of mVenus p27K−-transduced LCSC6 (left panel) and CSC7 (right panel) untreated and treated with NanoFEN at the IC50 dose (4,9 μM and 0,2 μM respectively) for 48 h, Gem 25 μM and OXA 10 μM. Values represent mean ± SD of three independent experiments. **P < 0.01; ***P < 0.001 from two-tailed t-test. d Schematic representation of stem-like factors (repressed in grey) emerged from RPPA analysis of lung CSCs treated as described in Fig. 4a. e Cytofluorimetric analysis of Aldefluor activity on LCSC6 treated with NanoFEN at IC50 dose and Gemcytabine 25 μM for 48 h. Values represent mean ± SD of three independent experiments. **P < 0.01; ***P < 0.001 from two-tailed t-test. f qRT-PCR analysis of indicated genes on whole tumor lysates derived from LCSC6 xenografts, untreated or treated with NanoFEN 30 mg/Kg/week and Cis 3 mg/Kg/week and Gem 60 mg/Kg/week (Cis + Gem). Values represent mean ± SD of three independent experiments. *P < 0.05; **P < 0.01 from two-tailed t-test. g Flow cytometry analysis of TOP-GFP.mc-transduced CSC7, untreated and treated with NanoFEN at the IC50 dose 0.2 μM and OXA 10 μM for 48 h. Values represent mean ± SD of three independent experiments. **P < 0.01 from two-tailed t-test. h Volume of xenografts derived from CSC10 transduced with TOP-GFP.mc untreated (black downward triangles) or treated with vehicle or treated NanoFEN 30 mg/Kg/week (green upward triangles) and OXA 5 mg/Kg/week plus 5FU 12.5 mg/Kg/week (red squares). Values represent mean ± SEM three independent experiments. *P < 0.05; **P < 0.01 from one-way ANOVA and Bonferroni post-test. i Total stem cell content in tumor xenografts obtained as in (h), relative to the experiment shown in Supplementary Fig. 3c. Values were calculated as tumor weight × percentage of TOP-GFP.mc positive cells/100. Values represent mean ± SD of three independent experiments. *P < 0.05; ***P < 0.001 from two-tailed t-test

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