Journal: Journal of Cellular and Molecular Medicine

Article Title: Targeting YAP in malignant pleural mesothelioma

doi: 10.1111/jcmm.13182

Figure Lengend Snippet: YAP activation and GTIIC reporter activity in cell lines. Five mesothelioma cell lines (211H, H2052, H2452, H290, MS‐1) and one normal mesothelial cell line (LP9) were measured by Western blotting and GTIIC reporter assay. ( A ) Western blot analysis of YAP and phospho‐YAP Ser127 (p‐YAP) in cell lines. ( B ) p‐YAP/YAP ratios were measured and are shown. Lower p‐YAP/YAP ratio indicates higher YAP activation status (* P < 0.05, *** P < 0.001, One‐way ANOVA and Scheffe multiple comparisons). ( C ) GTIIC reporter activity of the Hippo pathway in cell lines (**** P < 0.0001, One‐way anova and Scheffe multiple comparisons). ( D ) Cell viability analysis in 211H, H2052, H290, MS‐1, H2452 and LP9 cell lines after verteporfin treatment. IC 50 and genetic inactivation status are shown.

Article Snippet: Human mesothelioma cell lines 211H, H2052 and H2452 were purchased from American Type Culture Collections (ATCC, Manassas, VA, USA).

Techniques: Activation Assay, Activity Assay, Western Blot, Reporter Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Targeting YAP in malignant pleural mesothelioma

doi: 10.1111/jcmm.13182

Figure Lengend Snippet: Analysis of YAP protein level, GTIIC reporter activity, mRNA level of CTGF, cell invasion and tumoursphere formation in mesothelioma cells. ( A ) Western blotting analysis of YAP expression in H2052, H290 and 211H cells treated with the YAP inhibitor verteporfin. ( B ) A dose‐dependent decrease in GTIIC reporter activity of the Hippo pathway was analysed in H2052, H290 and 211H cells (** P < 0.01, **** P < 0.0001, One‐way anova , Scheffe multiple comparisons). ( C ) Decreased mRNA levels of CTGF and AREG, the downstream genes of the Hippo pathway, in H2052, H290 and 211H cells (* P < 0.05, ** P < 0.01, **** P < 0.0001, One‐way anova , Scheffe multiple comparisons). ( D ) Decrease in cell invasive ability after verteporfin treatment in H2052 cells. Images were taken under a 20 × objective lens. ( E ) Quantitative analysis of the number of cells that invaded the lower side of the membrane in each experimental group (** P < 0.01, *** P < 0.001, Student's t ‐test). ( F ) Decrease in sphere formation ability in H290 cells after verteporfin treatment. Images were taken under a 10 × objective lens. ( G ) Quantitative analysis of tumoursphere assay shows verteporfin treatment decreased tumoursphere formation ability in H290 cells (* P < 0.05, One‐way anova and Scheffe multiple comparisons).

Article Snippet: Human mesothelioma cell lines 211H, H2052 and H2452 were purchased from American Type Culture Collections (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Western Blot, Expressing, Membrane

Journal: Journal of Cellular and Molecular Medicine

Article Title: Targeting YAP in malignant pleural mesothelioma

doi: 10.1111/jcmm.13182

Figure Lengend Snippet: Analysis of YAP protein level, GTIIC reporter activity and mRNA level of YAP and its downstream genes after YAP inhibition by siRNAs. ( A ) Decreased YAP protein level after YAP siRNA‐1 and YAP siRNA‐2 treatment in H2052, H290 and 211H cells. ( B ) GTIIC reporter activity of the Hippo pathway after YAP inhibition by the two siRNAs was analysed in H2052, H290 and 211H cells (*** P < 0.001, **** P < 0.0001, one‐way anova and Scheffe multiple comparisons). ( C ) Decreased mRNA levels of CTGF and AREG, the downstream genes of the Hippo pathway, in H2052, H290 and 211H cells treated with the YAP siRNA‐1 (* P < 0.05, ** P < 0.01, *** P < 0.001, Student's t ‐test).

Article Snippet: Human mesothelioma cell lines 211H, H2052 and H2452 were purchased from American Type Culture Collections (ATCC, Manassas, VA, USA).

Techniques: Activity Assay, Inhibition

Journal: Journal of Cellular and Molecular Medicine

Article Title: Targeting YAP in malignant pleural mesothelioma

doi: 10.1111/jcmm.13182

Figure Lengend Snippet: Inhibition of RhoA/ROCK signaling suppresses GTIIC reporter activity and cell viability of mesothelioma cells. ( A ) GTIIC reporter activity of the Hippo pathway after RhoA, ROCK1 and ROCK2 siRNA was analysed in H2052 and 211H cells (* P < 0.05, ** P < 0.01, **** P < 0.0001, one‐way anova and Scheffe multiple comparisons). ( B ) GTIIC reporter activity of the Hippo pathway after different doses of ROCK inhibitor GSK269962A was analysed in H2052 and 211H cells (** P < 0.01, *** P < 0.001, **** P < 0.0001, one‐way anova and Scheffe multiple comparisons). ( C ) Cell viability analysis in 211H, H2052, H290, MS‐1, H2452 and LP9 cell lines, after GSK269962A treatment.

Article Snippet: Human mesothelioma cell lines 211H, H2052 and H2452 were purchased from American Type Culture Collections (ATCC, Manassas, VA, USA).

Techniques: Inhibition, Activity Assay

Journal: Cancers

Article Title: Exploring MicroRNA and Exosome Involvement in Malignant Pleural Mesothelioma Drug Response

doi: 10.3390/cancers14194784

Figure Lengend Snippet: Dose response curves depicting an enhanced MPM cell sensitivity to drug treatment with ( A ) cisplatin, ( B ) gemcitabine, ( C ) YM155 and ( D ) PND-1186, following transfection with various miRNAs (1 nM). The FAK inhibitor-resistant VMC23 cell line was used for all experiments involving PND-1186 treatment. The chemotherapy drug and survivin small molecule inhibitor-resistant MSTO-211H cell line was used for all experiments involving cisplatin, gemcitabine and YM155 treatment. All dose responses are shown with respect to the miRNA control mimic. Error bars represent the mean ± SD, as determined from three experimental replicates per tested drug concentration.

Article Snippet: Five MPM cell lines (H2052, H2452, H28, H226 and MSTO) and the immortalised mesothelial cell line, MeT-5A, were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Transfection, Control, Concentration Assay

Journal: Cancers

Article Title: Exploring MicroRNA and Exosome Involvement in Malignant Pleural Mesothelioma Drug Response

doi: 10.3390/cancers14194784

Figure Lengend Snippet: IC 50 values and fold change increase in MPM cell sensitivity to chemotherapy/small molecule inhibitor drug treatment following miRNA transfection pre-treatment of MPM cells.

Article Snippet: Five MPM cell lines (H2052, H2452, H28, H226 and MSTO) and the immortalised mesothelial cell line, MeT-5A, were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Transfection, Control

Journal: Cancers

Article Title: Exploring MicroRNA and Exosome Involvement in Malignant Pleural Mesothelioma Drug Response

doi: 10.3390/cancers14194784

Figure Lengend Snippet: Graphs depicting ( A ) MPM cell response to increasing concentrations of YM155 treatment; ( B ) quantified levels of exosome production in conditioned medium obtained from MPM cell cultures, with and without exosome inhibitor (GW4869), determined using an exosome prep kit; and ( C ) MSTO-211H response to increasing concentrations of YM155 following pre-treatment of MSTO-211H cells with the GW4896 exosome inhibitor, with respect to MSTO-211H cells not pre-treated with GW4896. Error bars represent the mean ± SD, as determined from three experimental replicates. Statistically significant differences in exosome production between MPM cells treated with and without exosome inhibitor (GW4869) are indicated with a single asterisk (*) for a p -value of ≤0.05 and with a double asterisk (**) for a p -value of ≤0.01.

Article Snippet: Five MPM cell lines (H2052, H2452, H28, H226 and MSTO) and the immortalised mesothelial cell line, MeT-5A, were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Cancers

Article Title: Exploring MicroRNA and Exosome Involvement in Malignant Pleural Mesothelioma Drug Response

doi: 10.3390/cancers14194784

Figure Lengend Snippet: Graphs depicting ( A ) mRNA expression levels corresponding to genes known to play a role in chemotherapy drug resistance, relative to the non-malignant MeT-5A control, as determined via qPCR analysis on a range of MPM cell lines; ( B ) mRNA levels corresponding to the ABCA6 and ABCA10 genes, relative to the non-malignant MeT-5A control, as determined via qPCR analysis on a range of MPM cell lines; MSTO-211H growth response to increasing concentrations of YM155 following silencing of the ( C ) ABCA6 and ( D ) ABCA10 genes. The dose response curves of the siRNA-treated cells are shown with respect to the untreated miRNA control mimic. Error bars represent the mean ± SD, as determined from three experimental replicates.

Article Snippet: Five MPM cell lines (H2052, H2452, H28, H226 and MSTO) and the immortalised mesothelial cell line, MeT-5A, were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Control

Journal: Oncology Letters

Article Title: Organic extract of Geodia cydonium induces cell cycle block in human mesothelioma cells

doi: 10.3892/ol.2022.13406

Figure Lengend Snippet: GEOCYDO treatment affects cell viability. Cell viability after treatment with various concentrations of GEOCYDO for 24 h in (A) MSTO or (B) Met-5A cells. (C) Cell viability after treatment of MSTO, NCI and Mes2 cells with 500 µg/ml GEOCYDO for different durations. Cells treated with vehicle only (0.1% DMSO) were used as a control. Data are presented as the mean ± standard deviation (n=3). ***P<0.005, ****P<0.001 vs. control. MSTO, MSTO-211H; NCI, NCI-H2452; Mes2, Ist-Mes2; GEOCYDO, Geodia cydonium extract.

Article Snippet: Human mesothelioma cell lines MSTO and NCI, and the human mesothelial cell line MeT-5A were grown in RPMI supplemented with 10% FBS (both from Euroclone SpA), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 IU/ml penicillin and 0.02 mg/ml streptomycin).

Techniques: Control, Standard Deviation

Journal: Oncology Letters

Article Title: Organic extract of Geodia cydonium induces cell cycle block in human mesothelioma cells

doi: 10.3892/ol.2022.13406

Figure Lengend Snippet: GEOCYDO treatment affects tumorigenic properties. (A) Colony formation assay on mesothelioma cell lines following GEOCYDO treatment. Representative plate images after crystal violet staining are shown. (B) Histograms of the average colony numbers. (C) Wound-healing assay. The wound closure rate was measured by detecting the closure distance at the time indicated in MSTO, NCI and Mes2 cells treated with 500 µg/ml GEOCYDO (+). Cells treated with vehicle only (0.1% DMSO) were used as a control (−). Representative micrographs under a phase contrast microscope are shown. Scale bar, 200 µm. (D) Quantification of wound gap distance. Data are presented as the mean ± standard deviation (n=3). ***P<0.005, ****P<0.001 vs. control. MSTO, MSTO-211H; NCI, NCI-H2452; Mes2, Ist-Mes2; GEOCYDO, Geodia cydonium extract.

Article Snippet: Human mesothelioma cell lines MSTO and NCI, and the human mesothelial cell line MeT-5A were grown in RPMI supplemented with 10% FBS (both from Euroclone SpA), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 IU/ml penicillin and 0.02 mg/ml streptomycin).

Techniques: Colony Assay, Staining, Wound Healing Assay, Control, Microscopy, Standard Deviation

Journal: Oncology Letters

Article Title: Organic extract of Geodia cydonium induces cell cycle block in human mesothelioma cells

doi: 10.3892/ol.2022.13406

Figure Lengend Snippet: Effect of GEOCYDO on cell cycle progression. (A) Distribution of MSTO cell population during the cell cycle after 16, 24 and 48 h of treatment with 500 µg/ml GEOCYDO (+) analyzed by flow cytometry. Cells treated with vehicle only (0.1% DMSO) were used as a control (−). (B) Histograms of the cell cycle distribution in MSTO, NCI and Mes2 cells. Data are presented as the mean ± standard deviation (n=3). MSTO, MSTO-211H; NCI, NCI-H2452; Mes2, Ist-Mes2; GEOCYDO, Geodia cydonium extract; P3, G 0 /G 1 cell population; P4, S cell population; P5, G 2 /M cell population.

Article Snippet: Human mesothelioma cell lines MSTO and NCI, and the human mesothelial cell line MeT-5A were grown in RPMI supplemented with 10% FBS (both from Euroclone SpA), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 IU/ml penicillin and 0.02 mg/ml streptomycin).

Techniques: Flow Cytometry, Control, Standard Deviation

Journal: Oncology Letters

Article Title: Organic extract of Geodia cydonium induces cell cycle block in human mesothelioma cells

doi: 10.3892/ol.2022.13406

Figure Lengend Snippet: GEOCYDO induces cell cycle arrest at G 0 /G 1 phase through modulation of cyclins and CDK inhibitors. Western blot analysis of the expression levels of cyclin E, cyclin A, cyclin B1, p21 and p27 16 h, 24 or 48 h after treatment with GEOCYDO. β-actin expression was used as a loading control. Actin 1 refers to the control for cyclin E, cyclin B1 and p21; Actin 2 refers to the control for cyclin A and p27; Actin 3 refers to the control for cyclin E, cyclin B1 and cyclin A (for NCI cells); Actin 4 refers to the control for p21; Actin 5 refers to the control for p27 and cyclin A (for MSTO and Mes2 cells); Actin 6 refers to the control for cyclin E, cyclin B1 and p21; Actin 7 refers to the control for p27 and cyclin A (for MSTO and Mes2 cells); Actin 8 refers to the control for cyclin A (for NCI cells). Histograms represent the relative expression levels relative to the control. All the controls were set at 100%. Data are presented as the mean ± standard deviation (n=3). *P<0.05, **P<0.01, ***P<0.005, ****P<0.001 vs. control. MSTO, MSTO-211H; NCI, NCI-H2452; Mes2, Ist-Mes2; GEOCYDO, Geodia cydonium extract.

Article Snippet: Human mesothelioma cell lines MSTO and NCI, and the human mesothelial cell line MeT-5A were grown in RPMI supplemented with 10% FBS (both from Euroclone SpA), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 IU/ml penicillin and 0.02 mg/ml streptomycin).

Techniques: Western Blot, Expressing, Control, Standard Deviation

Journal: Oncology Letters

Article Title: Organic extract of Geodia cydonium induces cell cycle block in human mesothelioma cells

doi: 10.3892/ol.2022.13406

Figure Lengend Snippet: Fraction C affects cell viability. (A) MSTO, NCI and Mes2 cells were treated with 200 mg/ml solid-phase extraction fractions B-E for 24 h. (B) MSTO, NCI and Mes2 cells were treated with 50–200 µg/ml fraction C for 24 h. Cells treated with vehicle only (0.1% DMSO) were used as a control. Data are presented as the mean ± SD of independent experiments (n=3). *P<0.05, **P<0.01, ****P<0.0001 vs. control. MSTO, MSTO-211H; NCI, NCI-H2452; Mes2, Ist-Mes2.

Article Snippet: Human mesothelioma cell lines MSTO and NCI, and the human mesothelial cell line MeT-5A were grown in RPMI supplemented with 10% FBS (both from Euroclone SpA), glutamine (2 mM), sodium pyruvate and antibiotics (0.02 IU/ml penicillin and 0.02 mg/ml streptomycin).

Techniques: Extraction, Control

Journal: Scientific Reports

Article Title: Impact of metallothionein-knockdown on cisplatin resistance in malignant pleural mesothelioma

doi: 10.1038/s41598-020-75807-x

Figure Lengend Snippet: Scores of metallothionein expression specified by immunohistochemical staining.

Article Snippet: MPM cell lines MSTO-211H (pemetrexed-sensitive, biphasic ) and NCI-H2052 (cisplatin-sensitive, sarcomatoid ) as well as the cell line NCI-H2452 ( BAP1 -mutant, cisplatin-resistant , epithelioid ) were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Thermo Fisher Scientific, Massachusetts), USA.

Techniques: Expressing, Immunohistochemical staining, Staining