Journal: Cell Communication and Signaling : CCS

Article Title: Establishment of a mouse hepatocellular carcinoma tumoroid panel recapitulating inter- and intra- heterogeneity for disease modelling and combinatorial drug discovery

doi: 10.1186/s12964-025-02391-w

Figure Lengend Snippet: Alb-R26 Met primary HCC cells are characterised by heterogeneous expression and phosphorylation levels of signals and by resistance to RTKi. ( A ) Western blot of the indicated signalling proteins. Actin or Tubulin were used for normalization. HCC13 cells were used as a positive control (34). Quantifications are reported in Fig. S5B. Full western blot and ponceau staining are shown in Fig. S10-12. ( B ) Dot plot reporting the percentage of viable cells after treatment (1, 3, 10 µM) with the indicated RTKi (cabozantinib, lenvatinib, sorafenib, and regorafenib). Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparison. Levels of significance: * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001

Article Snippet: The drugs used on cell cultures were: cabozantinib, lenvatinib, regorafenib, and sorafenib (1, 3, and 10μM; TargetMol: #T2586, #T0520, #T1792, #T0093L, respectively).

Techniques: Expressing, Phospho-proteomics, Western Blot, Positive Control, Staining, Comparison, Cell Viability Assay

Journal: Gastroenterology

Article Title: N6-Methyladenosine-Mediated Up-Regulation of FZD10 Regulates Liver Cancer Stem Cells' Properties and Lenvatinib Resistance Through WNT/β-Catenin and Hippo Signaling Pathways.

doi: 10.1053/j.gastro.2023.01.041

Figure Lengend Snippet: Figure 5. FZD10 determines the response of lenvatinib in HCC. (A) Kaplan-Meier analysis of overall survival in patients with HCC with and without lenvatinib therapy after recurrence. Left: all patients (P ¼ .077); middle: patients with low FZD10 expression (P ¼ .047); right: patients with high FZD10 expression (P ¼ .779). Median FZD10 expression was used as a cutoff. (B) Huh7/SNU398 FZD10 and control cells were treated with lenvatinib for 48 hours, and their cell survival curves were calculated. (C) Huh7 FZD10 and control cells were treated with lenvatinib for 10 days and their colony growth was examined. (D) Huh7/SNU398 FZD10 and control cells were treated with lenvatinib (10 mM) for 48 hours and their apoptosis was examined by flow cytometry. (E) PDOs derived from the primary HCCs with high FZD10 levels (patient nos. 5–7) or low FZD10 levels (patient nos. 8 and 9) were treated with lenvatinib for 7 days and their cell survival curves were calculated. (F) PDXs derived from primary HCCs with high FZD10 levels (patient nos. 5–7) or low FZD10 levels (patient nos. 8 and 9) were treated with lenvatinib (4 mg/kg) or saline for 4 weeks (n ¼ 5 for each group). The xenograft growth was monitored. (G) PDXs derived from indicated patients treated with saline or lenvatinib were subjected to Ki67 staining. Representative views were shown. Scale bar: 25 mm. (H) PDXs derived from indicated patients treated with saline or lenvatinib were subjected to Ki67 staining. The proportion of the Ki67-positive cells was quantified. All results are presented as mean ± SD and statistical significance was assessed using a 2-tailed Student t test. *P < .05.

Article Snippet: In order to establish lenvatinib-resistant cell lines, Huh7 and SNU398 cells were incubated with lenvatinib (Selleck, S1164) at a concentration just below their 50% inhibitory concentration, and increased by 0.2 mM/L per week for 6–7 months.

Techniques: Expressing, Control, Cytometry, Derivative Assay, Saline, Staining

Journal: Gastroenterology

Article Title: N6-Methyladenosine-Mediated Up-Regulation of FZD10 Regulates Liver Cancer Stem Cells' Properties and Lenvatinib Resistance Through WNT/β-Catenin and Hippo Signaling Pathways.

doi: 10.1053/j.gastro.2023.01.041

Figure Lengend Snippet: Figure 6. FZD10/b-catenin/c-Jun/ MEK/ERK axis determines lenva- tinib response in HCC. (A) Western blot analysis of indicated proteins in Huh7/SNU398 and Huh7LR/ SNU398LR cells. (B) Immuno- staining of FZD10, c-Jun, and p- ERK1/2 in matched human HCC taken before lenvatinib therapy (pre therapy) and after lenvatinib resistance (post relapse). (C) Huh7/SNU398 FZD10 and control cells treated with lenvatinib (2 mM) for 6 hours were subjected to Western blot analysis. (D) Huh7/ SNU398 FZD10 and control cells were transfected with sib-catenin or sic-Jun for 48 hours and then subjected to Western blot assay. (E) Huh7/SNU398 FZD10 and control cells were transfected with sib-catenin or sic-Jun. Cells were then treated with lenvatinib (10 mM) for 48 hours and their apoptosis was examined by means of flow cytometry. (F) Huh7 FZD10 and control cells were treated with FH535 (40 nM) or not for 48 hours and then subjected to Western blot assay. (G) Huh7/ SNU398 FZD10 and control cells was treated with FH535 (40 nM) or not. The cells were then treated with lenvatinib for 48 hours and cell survival curves were calcu- lated. (H) Huh7/SNU398 FZD10 and control cells were treated with FH535 (40 nM) or not. The cells were then treated with lenvatinib (10 mM) for 48 hours and their apoptosis was examined by flow cytometry. All results are pre- sented as the mean ± SD, and statistical significance was assessed using a 2-tailed Student t test. *P < .05.

Article Snippet: In order to establish lenvatinib-resistant cell lines, Huh7 and SNU398 cells were incubated with lenvatinib (Selleck, S1164) at a concentration just below their 50% inhibitory concentration, and increased by 0.2 mM/L per week for 6–7 months.

Techniques: Western Blot, Immunostaining, Control, Transfection, Cytometry

Journal: International journal of molecular sciences

Article Title: MicroRNA-142-3p Overcomes Drug Resistance in Hepatocellular Carcinoma by Targeting YES1 and TWF1.

doi: 10.3390/ijms26094161

Figure Lengend Snippet: Figure 1. Identification of miR-142-3p as a potential tumor suppressor miRNA in HCC. (A) Waterfall plot depicting the correlation between basal miRNA expression and TKI (sorafenib/lenvatinib) sensitivity in human cancer cell lines (data source: Cancer Cell Encyclopedia and Cancer Therapeutic Response Portal). (B) RNA-sequencing data of transcript per million (TPM) values of miR-142-3p expression in the sorafenib responder vs. non-responder group (BIOSTORM HCC cohort) from GSE109211 in the GEO datasets. Error bars = SD; *** p ≤0.001. (C) Kaplan–Meier plot of 166 HCC patients at different stages of the disease to determine the prognostic value of miR-142-3p expression. The p-value was calculated using the log-rank test; ** p = 0.0015. Abbreviations: SF: sorafenib; HR: hazard ratio.

Article Snippet: Cell viability was assessed at 72 h and 48 h post-treatment with lenvatinib (Selleck Chemicals, Houston, TX, USA; Cat# S5240) and dasatinib (Selleck Chemicals, Houston, TX, USA; Cat# S1021), respectively, using a CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega, Madison, WI, USA) according to the manufacturer’s guidelines.

Techniques: Expressing, Clinical Proteomics, RNA Sequencing

Journal: International journal of molecular sciences

Article Title: MicroRNA-142-3p Overcomes Drug Resistance in Hepatocellular Carcinoma by Targeting YES1 and TWF1.

doi: 10.3390/ijms26094161

Figure Lengend Snippet: Figure 7. Role of miR-142-3p and its gene targets in modulating TKI sensitivity in HCC cells. (A) SNU475 cells were reverse-transfected with 10 different doses of miR-142-3p mimic or miR-NC in 1/3 serial dilutions (highest dose 100nM), and at 48 h post-transfection were treated with lenvatinib with 10 doses in 1/2 dilution (highest dose 50µM). After 72 h treatment, cell viability was determined using cell titer assay. The combination index (CI) value was determined using the Chou–Talalay method. The IC50 values of lenvatinib alone and in combination with miR-142-3p are shown in graphs. (B) Incucyte proliferation assay of miR-142-3p ± lenvatinib. SNU475 cells were transfected

Article Snippet: Cell viability was assessed at 72 h and 48 h post-treatment with lenvatinib (Selleck Chemicals, Houston, TX, USA; Cat# S5240) and dasatinib (Selleck Chemicals, Houston, TX, USA; Cat# S1021), respectively, using a CellTiter 96 AQueous One Solution Cell Proliferation Assay (Promega, Madison, WI, USA) according to the manufacturer’s guidelines.

Techniques: Transfection, Titer Assay, Proliferation Assay

Journal: BMC Cancer

Article Title: Elevated expression of RAB3B plays important roles in chemoresistance and metastatic potential of hepatoma cells

doi: 10.1186/s12885-022-09370-1

Figure Lengend Snippet: Susceptibility of SK-HEP-1 derivative cells to anticancer drugs. The viability of SK-HEP-1 (white columns), RAB3B -KD (blue columns), and KD/pRAB3B (salmon columns) cells in the presence of anticancer drugs (5-fluorouracil, docetaxel, doxorubicin, irinotecan, SAHA, sorafenib, lenvatinib, regorafenib, and cabozantinib) was evaluated using the MTS assay. For each anticancer drug, the left and right three columns represent data for cells in the control and sphere inducing cultures, respectively. * P < 0.05 with Tukey–Kramer multiple comparison test

Article Snippet: Cells in culture medium were incubated with one of the following anticancer drugs, 10 mM 5-fluorouracil (5-FU, Sigma-Aldrich), 250 nM docetaxel (Sigma-Aldrich), 2 μM doxorubicin (Sigma-Aldrich), 200 μM irinotecan hydrochloride (Sigma-Aldrich), 2.5 μM suberoylanilide hydroxamic acid (SAHA, Cosmo Bio), 75 μM sorafenib tosylate (ChemScene, Monmouth Junction, NJ), 25 μM lenvatinib mesylate (Carbosynth, Berkshire, UK), 50 μM regorafenib (ChemScene), or 20 μM cabozantinib S-malate (ChemScene), 24 h at 37 °C in an atmosphere of 5% CO 2 in air.

Techniques: MTS Assay, Control, Comparison

Journal: Cell Death Discovery

Article Title: Sotorasib-impaired degradation of NEU1 contributes to cardiac injury by inhibiting AKT signaling

doi: 10.1038/s41420-025-02431-x

Figure Lengend Snippet: A CC-HEH-2 cells were treated with 0, 7.5, 15, 30, 60 μM sotorasib for 48 h; H9c2 cells were treated with 0, 7.5, 15, 30, 60 μM sotorasib for 48 h ( n = 3). Representative pictures. Scale bars: 50 μm. B CCC-HEH-2 cells were treated with 0, 3.75, 7.5, 15, 30, 60 μM sotorasib for 48 h; H9c2 cells were treated with 0, 3.75, 7.5, 15, 30, 60 μM sotorasib for 48 h. Survival fraction detection by SRB colorimetric assay ( n = 3). C CCC-HEH-2 cells were exposed to different concentrations of 0, 10, 15, or 20 μM sotorasib for 48 h. H9c2 cells were exposed to different concentrations of 0, 30, 45, or 60 μM sotorasib for 48 h. Expression levels of c-PARP protein in CCC-HEH-2 and H9c2 cells. D Male C57BL/6J mice were treated with 0.5% CMC-Na or sotorasib (300 mg/kg/d or 600 mg/kg/d) by gavage for 28 days ( n = 6). Representative image of IHC staining of cardiomyocytes c-CAPSP3 ( n = 3). Scale bars: 100 μm. E CCC-HEH-2 were treated with 20 μM sotorasib for 0, 12, 24, 36, and 48 h and exposed to different concentrations of 0, 10, 15, or 20 μM sotorasib for 48 h. The apoptosis rate of CCC-HEH-2 cells ( n = 3). F 20 μM sotorasib and/or 20 μM Z-VAD-FMK treated CCC-HEH-2 for 48 h. Expression level of c-PARP in CCC-HEH-2 cells ( n = 3). G CCC-HEH-2 were exposed to different concentrations of 0, 10, 15, or 20 μM sotorasib for 48 h. MMP changes in CCC-HEH-2 cells ( n = 3). H CCC-HEH-2 and were exposed to different concentrations of 0, 10, 15, or 20 μM sotorasib for 24 h. ATP levels in CCC-HEH-2 cells ( n = 3). I 20 μM sotorasib treatment of CCC-HEH-2 cells for 0, 6, 9, 12 and 24 h. ROS levels in CCC-HEH-2 cells ( n = 3). J Male C57BL/6J mice were treated with 0.5% CMC-Na or sotorasib (300 mg/kg/d or 600 mg/kg/d) by gavage for 28 days ( n = 6). Transmission electron microscopy representative images. scale bars: 2 μm. Data expressed as mean ± SD, ***, p < 0.0001, **, p < 0.01, *, p < 0.05 (vs. Vehicle), #, p < 0.05 (vs. sotorasib), &, p < 0.05 (vs. Z-VAD-FMK), ns, no significance.

Article Snippet: Materials: Sotorasib (T8684, Topscience, Shanghai, China), Z-VAD-FMK (T8541, Topscience, Shanghai, China), dexpanthenol (T1437, Topscience, Shanghai, China), D-PAC (T6480, Topscience, Shanghai, China), MG132 (M8699, Sigma-Aldrich).

Techniques: Colorimetric Assay, Expressing, Immunohistochemistry, Transmission Assay, Electron Microscopy

Journal: Current Issues in Molecular Biology

Article Title: NEAT1–SOD2 Axis Confers Sorafenib and Lenvatinib Resistance by Activating AKT in Liver Cancer Cell Lines

doi: 10.3390/cimb45020071

Figure Lengend Snippet: NEAT1 knockdown induces sorafenib and lenvatinib resistance. ( A ) Representative Western blot images for the indicated proteins. GAPDH is shown as an internal control. HLF and HuH6 cell lines were transduced with adenoviruses expressing nontargeting shRNA [shNT (N)] or NEAT1-specific shRNAs [shNEAT1a (Na) and shNEAT1b (Nb)] for 48 h. ( B ) Viabilities of HLF and HuH6 cells treated with sorafenib or lenvatinib at the concentrations indicated in the figure for 48 h relative to cells treated with dimethyl sulfoxide (DMSO; 100%). Cells were transduced with adenoviruses expressing shNT, shNEAT1a, and shNEAT1b 48 h before drug treatment. * p < 0.05 vs. shNT vs. shNEAT1a; # p < 0.05 shNT vs. shNEAT1b (Dunnett’s test; n = 4).

Article Snippet: Cells were treated with sorafenib (Adipogen Life Sciences, San Diego, CA, USA), lenvatinib (Toronto Research Chemicals, Toronto, ON, Canada), and capivasertib (Adooq Bioscience, Irvine, CA, USA) at the concentrations indicated in the figures, or DMSO as the control for 48 h in a 96-well plate.

Techniques: Knockdown, Western Blot, Control, Transduction, Expressing, shRNA

Journal: Current Issues in Molecular Biology

Article Title: NEAT1–SOD2 Axis Confers Sorafenib and Lenvatinib Resistance by Activating AKT in Liver Cancer Cell Lines

doi: 10.3390/cimb45020071

Figure Lengend Snippet: NEAT1v1 plays a role as a molecular switch of cell growth modality. ( A , C ) Representative Western blot images for the indicated proteins in control ( C ) or NEAT1v1-overexpressing (V1) cells. GAPDH is shown as an internal control. ( B , D ) Viabilities of control (CTRL) or NEAT1v1-overexpressing (NEAT1v1) cells treated with sorafenib ( B ), lenvatinib ( B ), or capivasertib ( D ) at the concentrations indicated in the figure for 48 h relative to cells treated with DMSO (100%). * p < 0.05 vs. CTRL (Student’s t -test; n = 4).

Article Snippet: Cells were treated with sorafenib (Adipogen Life Sciences, San Diego, CA, USA), lenvatinib (Toronto Research Chemicals, Toronto, ON, Canada), and capivasertib (Adooq Bioscience, Irvine, CA, USA) at the concentrations indicated in the figures, or DMSO as the control for 48 h in a 96-well plate.

Techniques: Western Blot, Control

Journal: Current Issues in Molecular Biology

Article Title: NEAT1–SOD2 Axis Confers Sorafenib and Lenvatinib Resistance by Activating AKT in Liver Cancer Cell Lines

doi: 10.3390/cimb45020071

Figure Lengend Snippet: NEAT1v1 regulates cell growth modality through SOD2. ( A ) Representative Western blot images for SOD2 expression in HLF and HuH6 cells transduced with adenoviruses expressing shNT (N), shNEAT1a (Na), or shNEAT1b (Nb) for 48 h. GAPDH is shown as an internal control. ( B ) Representative Western blot images for SOD2 expression in control ( C ) or NEAT1v1-overexpressing (V1) HLF and HuH6 cells. ( C , E ) Representative Western blot images for the indicated proteins in NEAT1v1-overexpressing cells transduced with adenoviruses expressing shNT (N) or SOD2-specific shRNAs [shSOD2a (Sa) and shSOD2b (Sb)] for 48 h. ( D , F ) Viabilities of cells treated with sorafenib ( D ), lenvatinib ( D ), or capivasertib ( F ) at the concentrations indicated in the figure for 48 h relative to cells treated with DMSO (100%). NEAT1v1-overexpressing cells were transduced with adenoviruses expressing shNT, shSOD2a, and shSOD2b 48 h before drug treatment. * p < 0.05 vs. shNT vs. shSOD2a; # p < 0.05 shNT vs. shSOD2b (Dunnett’s test; n = 4).

Article Snippet: Cells were treated with sorafenib (Adipogen Life Sciences, San Diego, CA, USA), lenvatinib (Toronto Research Chemicals, Toronto, ON, Canada), and capivasertib (Adooq Bioscience, Irvine, CA, USA) at the concentrations indicated in the figures, or DMSO as the control for 48 h in a 96-well plate.

Techniques: Western Blot, Expressing, Transduction, Control

Journal: Current Issues in Molecular Biology

Article Title: NEAT1–SOD2 Axis Confers Sorafenib and Lenvatinib Resistance by Activating AKT in Liver Cancer Cell Lines

doi: 10.3390/cimb45020071

Figure Lengend Snippet: NEAT1v1 activates the AKT pathway through SOD2, thereby conferring sorafenib and lenvatinib resistance in liver cancer cells, which are concomitantly sensitized to capivasertib. This result suggests that NEAT1v1 switches the growth modality of liver cancer cells from MEK/ERK-dependent to AKT-dependent mode via SOD2. Consistently, NEAT1 or SOD2 knockdown results in MEK/ERK activation, thereby sensitizing liver cancer cells to sorafenib and lenvatinib and conferring capivasertib resistance. NEAT1v1 or SOD2 knockdown also exacerbates ER stress; however, AKT is suppressed in an ER stress-independent manner.

Article Snippet: Cells were treated with sorafenib (Adipogen Life Sciences, San Diego, CA, USA), lenvatinib (Toronto Research Chemicals, Toronto, ON, Canada), and capivasertib (Adooq Bioscience, Irvine, CA, USA) at the concentrations indicated in the figures, or DMSO as the control for 48 h in a 96-well plate.

Techniques: Knockdown, Activation Assay