Journal: Frontiers in Oncology

Article Title: Inhibitory effect of the multi-target TKI, anlotinib, in 5-FU resistant colorectal cancer HCT-8/15 cells: down regulation of drug resistance-associated protein expression

doi: 10.3389/fonc.2026.1792045

Figure Lengend Snippet: Effect of anlotinib and 5-fluorouracil (5-FU) on colon cancer cell proliferation. (A) HCT-8/5-FU cells treated with 5-FU for 24 h and 48 h. IC50 (48 h) = 2246.5 ± 204.5 μM. (B) HCT-8/5-FU cells were treated with anlotinib for 24 h and 48 h. IC50 (24 h) = 53.69 ± 8.10 μM; IC50 (48 h) = 17.39 ± 1.98 μM. (C) HCT-15/5-FU cells treated with 5-FU for 24h and 48h. IC50 (48 h) = 18.49 ± 3.23 mM. (D) HCT-15/5-FU cells treated with anlotinib for 24 h and 48 h. IC50 (24 h) = 55.03 ± 3.44 μM; IC50 (48 h) = 28.83 ± 3.02 µM. Data are presented as the mean ± SD obtained from three independent experiments.

Article Snippet: HCT-8 cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA).

Techniques:

Journal: Frontiers in Oncology

Article Title: Inhibitory effect of the multi-target TKI, anlotinib, in 5-FU resistant colorectal cancer HCT-8/15 cells: down regulation of drug resistance-associated protein expression

doi: 10.3389/fonc.2026.1792045

Figure Lengend Snippet: Anlotinib reverses of 5-FU resistance in HCT-8/5-FU colon cancer cells. (A) HCT-8/5-FU cells were treated with anlotinib (at IC5 and IC10) and 5-FU at specified concentrations for 24 h. Cell viability was evaluated using the MTS assay. Data are presented as the mean ± SD. Doses below the IC10 are typically regarded as safe. In order to avoid cytotoxicity attributed to high doses rather than sensitization, we used IC10 in this experiment. (B) Colonies formed in three independent experiments were calculated from (C) . (C) Representative photos of HCT-8/5-FU colonies in the control, 6 µM anlotinib (IC10), 2 mM 5-FU (IC10), and combined treatment group. Following treatment with the indicated reagents, the number of colonies formed per dish was assessed. *p<0.05 represents the comparison between the experimental and the control group; #p<0.05 represents the comparison between the combination and single drug groups. Data represent three independent experiments.

Article Snippet: HCT-8 cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA).

Techniques: MTS Assay, Control, Comparison

Journal: Frontiers in Oncology

Article Title: Inhibitory effect of the multi-target TKI, anlotinib, in 5-FU resistant colorectal cancer HCT-8/15 cells: down regulation of drug resistance-associated protein expression

doi: 10.3389/fonc.2026.1792045

Figure Lengend Snippet: Effects of anlotinib on the cell cycle. (A, B) The effect of different anlotinib concentrations (0, 5, 10, 20 µM, in (a–d) respectively) on HCT-8/5-FU cells. (C, D) HCT-15/5-FU cells assayed as in (A, B) , (e–h) . Anlotinib reduced the number of cells in the S phase while increasing those in the G0/G1 phase (p<0.05). Concentrations of 20 µM and 10 µM had the greatest effect in HCT-15/5-FU and HCT-8/5-FU cells, respectively. Data are presented as the mean ± SD of three experiments. *p<0.05 represents the comparison between the experimental and control groups. Anlotinib suppressed the viability of HCT-8/FU cells even at 5 µM, while no effect was observed in HCT-15/FU cells. Therefore, 5 µM was not included as a treatment for HCT-15/FU cells. Meanwhile, 80 µM had the strongest cytotoxic effect. To avoid excess cell death, this dose was not selected.

Article Snippet: HCT-8 cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA).

Techniques: Comparison, Control

Journal: Frontiers in Oncology

Article Title: Inhibitory effect of the multi-target TKI, anlotinib, in 5-FU resistant colorectal cancer HCT-8/15 cells: down regulation of drug resistance-associated protein expression

doi: 10.3389/fonc.2026.1792045

Figure Lengend Snippet: Effect of anlotinib on drug resistance-associated protein expression and AKT signaling in 5-FU-resistant colorectal cancer cells. HCT-8/5-FU and HCT-15/5-FU cells were treated with increasing concentrations of anlotinib (IC5, IC10, and higher doses) for 24 h. Protein levels of MDR1, MRP1, phosphorylated AKT (p-AKT), and total AKT were analyzed by immunoblotting; GAPDH was used as a loading control.

Article Snippet: HCT-8 cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, USA).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: Efficacy of an XPO1 inhibitor in combination with irinotecan in a preclinical colorectal cancer model

doi: 10.3389/fonc.2026.1721685

Figure Lengend Snippet: In vitro evaluation of sequential dosing of SN38 and/or eltanexor in CRC cell lines (HCT8, HCT116, LS1034, and HCT15). (A) Dosing strategies for in vitro viability assay. Cells were incubated with SN38 [10 nM] for the first 24 hours. Cells were then washed with PBS and incubated with eltanexor [100 nM] for an additional 48 hours. (B) Cell viability % measured by CellTiter Glo 2.0 and (C) heatmaps of the 4 CRC cell lines treated sequentially with SN38 [0 - 30 nM] followed by eltanexor [0 -100 μM]. Bliss synergy score was analyzed using SynergyFinder.

Article Snippet: Human CRC cell lines HCT8 (RRID: CVCL_2478), HCT15 (RRID: CVCL_0292), HCT116 (RRID: CVCL_0291), and LS1034 (RRID: CVCL_1382) were purchased from American Type Culture Collection (ATCC) (Manassas, VA; ).

Techniques: In Vitro, Viability Assay, Incubation

Journal: Frontiers in Oncology

Article Title: Efficacy of an XPO1 inhibitor in combination with irinotecan in a preclinical colorectal cancer model

doi: 10.3389/fonc.2026.1721685

Figure Lengend Snippet: (A) A Western blot analysis of RAD51, p53, p-H2A.X and MSH2 in CRC cell lines HCT8, HCT116, LS1034, and HCT15 following sequential treatment of SN38 and/or eltanexor. (B) Densitometry analysis total cell protein. Cells were exposed to SN38 (10 nM) or vehicle for 6hr. (C) Cells were then washed with PBS and incubated with or without the presence of eltanexor (1 uM) for an additional 24 hr. Nuclear/Cytoplasmic. (D) Densitometry analysis of nuclear/cytoplasmic proteins.

Article Snippet: Human CRC cell lines HCT8 (RRID: CVCL_2478), HCT15 (RRID: CVCL_0292), HCT116 (RRID: CVCL_0291), and LS1034 (RRID: CVCL_1382) were purchased from American Type Culture Collection (ATCC) (Manassas, VA; ).

Techniques: Western Blot, Incubation

Journal: Frontiers in Oncology

Article Title: Efficacy of an XPO1 inhibitor in combination with irinotecan in a preclinical colorectal cancer model

doi: 10.3389/fonc.2026.1721685

Figure Lengend Snippet: Immunocytochemistry in HCT8 and HCT 116 cell lines. Cells were first treated with SN38 (10 nM) in time series (0 hr, 2 hr, 4 hr, and 6 hr). Cells were washed with PBS and then treated with eltanexor (1 uM) for 48 hours. Cells were fixed and then stained with p53 and p21 for cell cycle arrest or stained with P-H2A.X for double stained DNA breaks. (A) Schematic illustration of dosing strategy, (B) immunostaining in HCT8 cell line and (C) immunostaining in HCT116 cell line.

Article Snippet: Human CRC cell lines HCT8 (RRID: CVCL_2478), HCT15 (RRID: CVCL_0292), HCT116 (RRID: CVCL_0291), and LS1034 (RRID: CVCL_1382) were purchased from American Type Culture Collection (ATCC) (Manassas, VA; ).

Techniques: Immunocytochemistry, Staining, Immunostaining