Skip to main content
. 2022 Jul 22;12:942341. doi: 10.3389/fonc.2022.942341

Figure 4.

Figure 4

Abemaciclib combined with lenvatinib reduces 3D cell growth, inhibits cell migration, and induces G1 cell cycle arrest associated with senescence. (A) The growth of spheroids from HUH7 was analyzed after 6 or 12 days of treatment with 0.1 μM A and 0.5 μM L alone or in combination. The data are expressed as percent of spheroid growth versus control. Representative images of spheroids after 12 days of culture are shown. ***p<0.001 vs C; #p<0.05 vs A; §p<0.05 vs L. (B) SNU398 cells were treated with 0.2 μM A or 2 μM L as single treatment or in combination for 20h. Migrated cells were then counted. Data are expressed as percent versus control. Representative fields of migration are shown (magnification of 4×). ***p<0.001 vs C; ###p<0.001 vs A; §§§p<0.001 vs L. (C) HUH7, SNU398, and HepG2 cells were untreated (C) or treated with 0.1, 0.2 μM and 0.05 μM A and 0.5, 2 or 6.5 μM L for HUH7, SNU398 and HepG2, respectively, as single treatment or in combination for 24h. After 24h the cells were stained with propidium iodide and cell-cycle-phase distribution was determined by flow cytometry. Data are expressed as percentage of cells in each cell-cycle phase. **p<0.01, ***p<0.001 vs C; #p<0.05, ##p<0.01 vs A; §§§p<0.001 vs L. (D) HUH7 cells were treated with 0.1 μM A and 0.5 L alone or in combination for 6 days. Histograms represent the percentage of senescent cells positive for SA-β-Gal expression. ***p<0.001 vs C; ###p<0.001 vs A; §§§p<0.001 vs L. (E) HUH7 was treated with 0.1 μM A and 0.5 L alone or in combination for 3 days. The cells were lysed and the expression of the indicated proteins was evaluated by Western blot analysis. (F) HUH7 were treated with 0.1 μM A and 0.5 L alone or in combination. After 6 days, the cells were incubated with 1 μM CM-H2DCFDA probe and analyzed by flow cytometry. #p<0.05 vs A; §p<0.05 vs L. Experiments in A-E are representative of two independent experiments. Data in (B, C) are mean values ± SD of two independent experiments. Data in D are mean values ± SD of three independent experiments. Data in F are mean values ± SD of three independent experiments.