Journal: Frontiers in Endocrinology

Article Title: Identification of the enhancer RNAs related to tumorgenesis of pituitary neuroendocrine tumors

doi: 10.3389/fendo.2023.1149997

Figure Lengend Snippet: The effect of four molecular inhibitors on GH3 cell line in vitro (A) and the effect of ciclopirox on primary GH-secreting pituitary adenoma cells in vitro (B) . Cell survival was determined by CellTiter-Glo luminescent cell viability assay. The effect of ciclopirox in vivo . Representative images (C) , tumor growth curve (D) , mass weight (E) , representative images of HE, ki67 and PCNA staining on tumor section of mice indicated in (F) , and body weight (G) of tumors derived from athymic tumor-bearing mice treated with PBS or ciclopirox. *p<0.05; ***p<0.001.

Article Snippet: IHC was performed by blocking the rehydrated tissue sections using bovine serum overnight at 4°C, and subsequently applying the Anti-Ki67 antibody (Abcam, Cat. no. ab15580) and Anti-PCNA antibody (Abcam, Cat. no. ab92729).

Techniques: In Vitro, Cell Viability Assay, In Vivo, Staining, Derivative Assay

Journal: PLOS ONE

Article Title: Cytosolic EpCAM cooperates with H-Ras to regulate epithelial to mesenchymal transition through ZEB1

doi: 10.1371/journal.pone.0285707

Figure Lengend Snippet: A-B. Phase contrast image of MCF10A-Ras-EpCAM-L240A cells cultured until sub-confluent and treated for 36-h with Veh. (DMSO), MEKi (Ci-1040, 20nm), JNKi (SP600125, 1 μM), P38 MAKPi (SB203580, 100 nM), AKti (LY294002, 2 μM), NF-kBi (BAY117082, 10 μM), dasatinib (100 nm), doxorubicin (2 μM), etoposide (5 μM), paclitaxol (100 nm), and quercetin (5 μM) to assay drug sensitivity. B. Efficacy of inhibitors used were analyzed by western blotting. Cells were harvested after 36h of drugs treatment, lysates were immunoblotted to common pathways, ERK1/2, JNK, P38, AKT, NF-kB and SRC. C. The cell proliferation assay was performed after 72 of plating using CellTiter-Glo Luminescent Cell Viability Assay. D. Cell viability was determined using MTT assay (see methods). Results shows paclitaxel and quercetin reduced cell growth significantly. E. MCF10A-Ras-EpCAM-L240A cells cultured until sub-confluent and treated for 36-h with Vehicle, Doxorubicin, Etoposide, Paclitaxel, and quercetin. Cell lysates were subjected for immunoblotting to assay cleaved caspase-3 and PARP.

Article Snippet: The MAPK inhibitors CI-1040, SB203580, and SP600125 and AKT inhibitor LY29004 were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Cell Culture, Western Blot, Proliferation Assay, Cell Viability Assay, MTT Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro antineoplastic effects of brivaracetam and lacosamide on human glioma cells

doi: 10.1186/s13046-017-0546-9

Figure Lengend Snippet: Brivaracetam and lacosamide treatments exerted an anti-proliferative effect and a reduction in migration ability of glioma cells . a Distribution of T98G cells in the different phases of the cell cycle upon 24 h BRV or LCM treatments (IC20). Data are expressed as percentage of cell in a specific phase (G0/G1, S, G2/M) and refers to at least four independent experiments. Statistical evaluation was performed by the student’s t-test. Histogram bars represent mean ± standard deviation of at least three independent replicates. b - c - d Western-blot analysis of cyclin A, cyclin E, p53 and p21 upon treatments with BRV or LCM at IC20 in T98G a U87MG b SW1783 c cells. e Transwell migration assay in U87MG cells upon BRV or LCM treatments. (IC20). f Western-blot analysis of EGFR and N-cadherin proteins expression levels in U87MG cells upon BRV or LCM treatments (IC20)

Article Snippet: The human glioma cell lines U87MG, SW1483 and T98G and human fibroblasts were purchased from ATCC (LGC Standards S.r.l.).

Techniques: Migration, Standard Deviation, Western Blot, Transwell Migration Assay, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro antineoplastic effects of brivaracetam and lacosamide on human glioma cells

doi: 10.1186/s13046-017-0546-9

Figure Lengend Snippet: Brivaracetam and lacosamide treatments modulate microRNAs expression in U87MG glioma cells. a Heat map of the identified signature of 37 microRNAs differentially expressed in U87MG cells treated with BRV at IC20 at the indicated time points. b Heat map of the identified signature of 30 microRNAs differentially expressed in U87MG cells treated with LCM at IC20 at the indicated time points. c Supervised statistical test of the significance level of the difference between signal distributions of miR-107 and miR-195-5p in U87MG cells treated with LCM or BRV versus the control (untreated cells). d - e - f qRT-PCR validation of miR-107 and miR-195-5p in U87MG, SW1783 and T98G upon IC20 LCM or BRV treatments at the indicated time points

Article Snippet: The human glioma cell lines U87MG, SW1483 and T98G and human fibroblasts were purchased from ATCC (LGC Standards S.r.l.).

Techniques: Expressing, Control, Quantitative RT-PCR, Biomarker Discovery

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro antineoplastic effects of brivaracetam and lacosamide on human glioma cells

doi: 10.1186/s13046-017-0546-9

Figure Lengend Snippet: Brivaracetam and lacosamide treatments induce epigenetic modification on miR-107 and miR-195 regulatory regions. a - b qRT-PCR of miR-107 a and miR-195-5p b precursors (pre-miRNAs) in U87MG cells upon IC20 LCM or BRV treatments. c Schematic representation of miR-107 gene locus: two different regions of host gene (PANK1) promoter have been analyzed in ChIP assays (P1 and P2). d - e ChIP analysis of acetylated histone H4 and methylated histone H3 occupancy on miR-107 regulatory regions in U87MG cells upon treatment with BRV or LCM at IC20. f Schematic representation of miR-195-5p gene locus: two different regions of host gene (MIR497HG) promoter have been analyzed in ChIP assays (P1 and P2). g - h ChIP analysis of acetylated histone H4 and methylated histone H3 occupancy on miR-195-5p regulatory regions in U87MG cells upon treatment with BRV or LCM at IC20

Article Snippet: The human glioma cell lines U87MG, SW1483 and T98G and human fibroblasts were purchased from ATCC (LGC Standards S.r.l.).

Techniques: Modification, Quantitative RT-PCR, Methylation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro antineoplastic effects of brivaracetam and lacosamide on human glioma cells

doi: 10.1186/s13046-017-0546-9

Figure Lengend Snippet: Brivaracetam and lacosamide treatments exert their anti-proliferative effect in part trough miR-195-5p. a - b Proliferation assay ( a ) and viability assay b in U87MG cells transfected with miR-195-5p mimic or control. Cells were collected and counted at the indicated time points. c Distribution of U87MG cells in the different phases of the cell cycle. Data are expressed as percentage of cell in a specific phase (G0/G1, S, G2/M) and refers to at least four independent experiments. Statistical evaluation was performed by the student’s t-test. Histogram bars represent mean ± standard deviation of at least three independent replicates. d Western-blot analysis of cyclin A, cyclin E and p21 protein expression levels in U87MG cells upon miR-195-5p over-expression. e Viability assay in U87MG cells transfected with miR-195-5p inhibitor or control and treated with BRV or LCM at IC20. f Western-blot analysis of cyclin A, cyclin E and p21 protein expression levels in U87MG cells upon miR-195-5p depletion with miR-195-5p inhibitor (inh miR-195-5p). g Distribution of U87MG cells in the different phases of the cell cycle upon miR-195 depletion and treatments wit BRV or LCM at IC20. Data are expressed as percentage of cell in a specific phase (G0/G1, S, G2/M) and refers to at least four independent experiments. Statistical evaluation was performed by the student’s t-test. Histogram bars represent mean ± standard deviation of at least three independent replicates. h Western-blot analysis of cyclin A, cyclin E and p21 protein expression levels in U87MG cells upon miR-195-5p depletion with miR-195-5p inhibitor (inh miR-195-5p) and treatments wit BRV or LCM at IC20. (* = p val < 0.05)

Article Snippet: The human glioma cell lines U87MG, SW1483 and T98G and human fibroblasts were purchased from ATCC (LGC Standards S.r.l.).

Techniques: Proliferation Assay, Viability Assay, Transfection, Control, Standard Deviation, Western Blot, Expressing, Over Expression

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: In vitro antineoplastic effects of brivaracetam and lacosamide on human glioma cells

doi: 10.1186/s13046-017-0546-9

Figure Lengend Snippet: Brivaracetam and lacosamide treatments inhibit glioma cells migration in part trough miR-107. a Transwell migration assay in U87MG cells upon miR-107 exogenous expression (* = p val < 0.05). b - c Western-blot analysis of EGFR and N-cadherin proteins expression levels in U87MG cells upon miR-107 over-expression with miR-107 mimic (miR-107) ( b ) or depletion with miR-107 inhibitor (inh miR-107) ( c ). d Transwell migration assay in U87MG cells upon miR-107 depletion with miR-107 inhibitor (inh miR-107)

Article Snippet: The human glioma cell lines U87MG, SW1483 and T98G and human fibroblasts were purchased from ATCC (LGC Standards S.r.l.).

Techniques: Migration, Transwell Migration Assay, Expressing, Western Blot, Over Expression

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: α-hederin inhibited the proliferation of HCC cell lines. (A) HepG2, SMMC-7721 or Huh-7 cells were treated with the indicated concentrations of α-hederin for 12, 24 or 48 hours, followed by MTT cell viability assay. (B) Cell cycle analysis of HepG2 and SMMC-7721 cells treated with α-hederin at low (10 µM) or high (20 µM) concentration for 24 hours. (C) Representative images and analysis of EdU staining in HepG2 and SMMC-7721 cells treated with α-hederin at low (10 µM) or high (20 µM) concentration for 24 hours. (D) Representative images and quantitative analysis of colony numbers of HepG2 and SMMC-7721 cells treated with α-hederin at low (10 µM) or high (20 µM) concentration for 24 hours. Significance: * p < 0.05 versus control, ** p < 0.01 versus control.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: Viability Assay, Cell Cycle Assay, Concentration Assay, Staining, Control

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: α-hederin promoted apoptosis of HCC cell lines in vitro . (A) Analysis of apoptosis in HepG2 and SMMC-7721 cells treated with α-hederin for 24 hours by flow cytometry. (B) The protein levels of Bax, Bcl-2, caspase 3, and cleaved caspase 3 were detected by Western blot in HepG2 and SMMC-7721 cells treated with the indicated concentrations of α-hederin for 24 hours. (C) Analysis of genes associated with cell proliferation and apoptosis in HepG2 and SMMC-7721 cells treated with α-hederin for 24 hours. Significance: * p < 0.05; ** p < 0.01 versus control.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: In Vitro, Flow Cytometry, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: α-hederin inhibited YAP activation via upregulating the activation of Hippo signaling pathway. (A) The protein and mRNA and levels of YAP in HepG2, SMMC-7721, Huh-7 cells. (B) HepG2 and SMMC-7721 cells were treated with α-hederin at low (10 µM) or high (20 µM) concentration for 24 h, and then the proteins involved in Hippo-YAP signaling pathway were analyzed by Western blot, followed by densitometric quantification. (C) The mRNA levels of Mst1, Lats1, YAP in HepG2 and SMMC-7721 cells treated with α-hederin for 24 hours were measured by RT-qPCR. (D) Immunofluorescence analysis of YAP in HepG2 and SMMC-7721 cells. (E) The nuclear levels of YAP in HepG2 and SMMC-7721 cells treated with α-hederin at low (10 µM) or high (20 µM) were analyzed by Western blot. Significance: * p < 0.05 versus control, ** p < 0.01 versus control.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: Activation Assay, Concentration Assay, Western Blot, Quantitative RT-PCR, Immunofluorescence, Control

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: Inhibition of Mst1 activation reversed the effect of α-hederin on hepatoma cell proliferation. (A) Western blot analysis of Mst1 and Lats1 expression in HepG2 and SMMC-7721 cells treated with XMU-MP-1 for 3 h or 6 h. (B) Flow cytometric analysis of apoptotic HepG2 and SMMC-7721 cells after α-hederin or α-hederin/XMU-MP-1 treatment. (C) Flow cytometric analysis of G1- and G2/M-phase subpopulations in HepG2 and SMMC-7721 cells treated with α-hederin or α-hederin/XMU-MP-1. (D) Cell proliferation assay using EDU labeling in HepG2 and SMMC-7721 cells treated with α-hederin or α-hederin/XMU-MP-1. Significance: ** p < 0.01 versus control, #p<0.05 versus High concentration, ## p < 0.01 versus High concentration.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Proliferation Assay, Labeling, Control, Concentration Assay

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: α-hederin-induced Mst1 upregulation was responsible to the inhibition of YAP signaling pathway. (A) The expression of proteins involved in Hippo-YAP signaling pathway in HepG2 and SMMC-7721 cells that treated with α-hederin or α-hederin/XMU-MP-1 was analyzed by Western blot. (B) The expression of Hippo-YAP signaling target genes in HepG2 and SMMC-7721 cells that treated with α-hederin or α-hederin/XMU-MP-1 was analyzed by RT-qPCR. (C) Analysis of genes associated with cell proliferation and apoptosis in HepG2 and SMMC-7721 cells treated with α-hederin or α-hederin/XMU-MP-1. (D) Representative immunofluorescence images of YAP in HepG2 and SMMC-7721 cells with the indicated treatment. Significance: * p < 0.05 versus control, ** p < 0.01 versus control, # p < 0.05 versus High concentration, ## p < 0.01 versus High concentration.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: Inhibition, Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Control, Concentration Assay

Journal: Frontiers in Oncology

Article Title: α-Hederin Inhibits the Proliferation of Hepatocellular Carcinoma Cells via Hippo-Yes-Associated Protein Signaling Pathway

doi: 10.3389/fonc.2022.839603

Figure Lengend Snippet: α-hederin attenuated xenograft HCC tumor growth in vivo through inhibiting YAP signaling pathway. (A) In vivo imaging of the HepG2 cell xenograft model mice. (B) Representative images of tumors collected from HepG2 cell xenograft model mice treated with PBS, α-hederin and DDP. (C) Weight of xenograft tumors harvested from mice. (D) Kidney index (kidney weight/bodyweight). (E) Tumor volumes were measured at different time points. (F) The ratio of tumor weight/body weight. (G) Mice body weight subtracted tumor at the end point. (H) HE staining of representative tumor tissues from different groups. (I) Immunohistochemistry analysis of YAP in mouse tumor tissues. (J) Immunoblot analysis of Hippo-YAP signaling pathway-related proteins and apoptosis-related proteins in the tumor tissues. (K) TUNEL staining of tumor tissues. Significance: * p < 0.05 versus control, ** p < 0.01 versus control.

Article Snippet: The human SMMC-7721, HepG2, and Huh-7 HCC cell lines were purchased from ATCC.

Techniques: In Vivo, In Vivo Imaging, Staining, Immunohistochemistry, Western Blot, TUNEL Assay, Control

Journal: Journal of biomedical materials research. Part A

Article Title: In vitro evaluation of bone cell response to novel 3D-printable nanocomposite biomaterials for bone reconstruction.

doi: 10.1002/jbm.a.37719

Figure Lengend Snippet: FIGURE 5 Differentiated MC3T3-E1 osteoblast cell proliferation on mSLA-printed nanocomposite discs. SP10 (10% nanohydroxyapatite [nHA]), mSP10 (methacrylated acrylated epoxidized soybean oil [mAESO] and 10% nHA), SP30 (30% nHA), mSP30 (mAESO and 30% nHA), and hydroxyapatite (HA) (control). Proliferation of dMC3T3-OB cells as measured by XTT assay. Data are means ± SD (n = 8 for nanocomposite discs and n = 5 for HA discs, from four experiments), (statistically significant, *p ≤.05, **p ≤.01, ***p ≤.001).

Article Snippet: Cell proliferation after 1, 3, and 7 days of culture was characterized using XTT (sodium 30-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis (4-methoxy6-nitro) benzene sulfonic acid hydrate; TACS XTT cell proliferation kit, R&D Systems, Minneapolis, USA).

Techniques: Control, XTT Assay