Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Characterization of Pttg2-depleted HCT116 cells. ( a ) Schematic diagram showing the sequences and location of shPttg1 and shPttg2. Multiple sequence alignments were made using the Clustal W software ( www.clustal.org ). ( b ) Relative quantification of Pttg1 and Pttg2 mRNA levels in Pttg1 and Pttg2-silenced HCT116 wild-type and HCT116 pttg1 −/− cells by real-time PCR. Expression of each gene is shown as mean±S.E.M. of three independent experiments. Data are shown as 2 −ΔCT normalized to HPRT1. ( c ) Analysis of PTTG1 and PTTG2 protein levels 72 h after shRNA-Pttg1 or shRNA-Pttg2 treatment in HCT116 wild-type and HCT116 pttg1 −/− cells. β -Actin was used as loading control. Densitometric analysis for western blots of PTTG1 and PTTG2 protein expression (% of control) in control and shRNA-treated (shPttg1 or shPttg2) wild-type and Pttg1 −/− HCT116 cells

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Sequencing, Software, Quantitative Proteomics, Real-time Polymerase Chain Reaction, Expressing, shRNA, Control, Western Blot

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Pttg1 and Pttg2 show different biochemical properties. ( a ) PTTG2 does not interact with Separase. Extracts of HEK293T cells expressing N-terminally Flag-tagged separase, C-terminally Myc-tagged PTTG2, or both were used for immunoprecipitation with anti-Myc antibodies. The immunoprecipitates were analyzed by immunoblotting with anti-FLAG and anti-Myc antibodies. The asterisks indicate the immunoglobulin light chains. As positive control, the interaction between PTTG1 and separase was tested using Flag-separase and Myc-tagged PTTG1. ( b ) PTTG2 lacks transactivation capacity. The ORF of Pttg2 was fused to the GAL4 DNA-binding domain (pGAL4-DBD-Pttg2) and tested for transcription activation in HEK293T cells cotransfected with a GAL4 site-dependent reporter plasmid driving expression of the luciferase gene (pGAL4-4RE-Luc). pGAL4-PTTG1 and pGAL4-VP16 were used as positive controls. Luciferase values from at least three independent transfections are plotted as fold activation over control

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Expressing, Immunoprecipitation, Western Blot, Positive Control, Binding Assay, Activation Assay, Plasmid Preparation, Luciferase, Transfection, Control

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Silencing of Pttg2 impairs cell adhesion and proliferative capacity of HCT116 cells. ( a ) Phase-contrast images ( × 10 objective) of Pttg2-silenced HCT116 cells 72 h (upper panels) and 1 week post-infection (PI) (lower panels). Inserts show close-up views of representative cell culture areas. Pttg2-depleted cells show rounder morphology than control cells, indicating a defect in cell adhesion properties. ( b ) Spheroid formation of Pttg2 and Pttg1-silenced HCT116 (wt) cells in suspension. Seventy-two hours post-infection, cells were transferred to poly-HEMA-coated plates at 6 × 10 4 cells/ml. After 48 h in suspension, cells were photographed at 10 × . Control and shPttg1-treated cells formed dense spheroids whereas shPttg2-silenced cells adhere to each other very weakly. Representative areas of each culture were selected. ( c ) Percentage of BrdU-positive cells after shRNA-Pttg2 treatment. HCT116 cells were infected and assayed for BrdU incorporation 72 h post-infection. Bars represent the mean±S.E.M. ( d ) Analysis of mitotic index in Pttg2-depleted HCT116 cells. Cells were treated with shRNA-Pttg2 lentivirus and incubated with nocodazole 1 μ M 72 h post-infection, for 24 h. Percentage of H3P-positive (mitotic) cells was measured by flow cytometry. Bars represent the mean±S.E.M. ( e ) Percentage of cell death following shRNA Pttg2 or shRNA Pttg1 treatment of HCT116 cells growing under adherent or suspension conditions. Seventy-two hours post-infection, cells were transferred onto control or poly-HEMA-coated (PH) plates in the absence or presence of methylcellulose (MC) and the levels of apoptosis was determined 48 h later by measuring the percentage of cells containing a subG1 DNA content by flow cytometry. Results represent the means of three independent experiments±S.E.M. ( f ) p21 induction in Pttg2-depleted HCT116 cells. HCT116 cells treated with shRNA-Pttg2 lentivirus for 72 h were harvested and the levels of p21 determined by immunoblotting using specific p21 antibodies. β -Actin was used as loading control

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Infection, Cell Culture, Control, Suspension, shRNA, BrdU Incorporation Assay, Incubation, Flow Cytometry, Western Blot

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Defective adhesion in Pttg2-depleted HCT116 cells results in p53- and p21-dependent apoptosis. ( a ) Phase-contrast images of Pttg2-silenced HCT116 cells after z-VAD treatment. A round morphology was still observed in Pttg2-depleted cells, suggesting a defective cell adhesion. ( b ) Apoptosis can be rescued in Pttg2-silenced HCT116 cells after caspase inhibition treatment. The percentage of apoptotic cells (subG1) after PTTG2 depletion was drastically reduced after treatment with the caspase inhibitor z-VAD. Bars represent the mean±S.E.M. ( c ) Levels of apoptotic indicators in Pttg2-silenced HCT116 cells. Cells were harvested after 72 h post-infection and assayed for p53, p21, BIM and cleaved-Capase 3 levels by western blot using specific antibodies. β -Actin was used as loading control. ( d , e ) Role of p53 in Pttg2-dependent anoikis. ( d ) Phase-contrast images of Pttg2-depleted HCT116 p53 −/− cells. The absence of PTTG2 in HCT116 p53 −/− results in cell rounding similar to wild-type HCT116 cells. ( e ) FACS analysis of Pttg2-depleted HCT116 p53 −/− cells. In the absence of PTTG2, the percentage of apoptotic cells (subG1) was partially reduced in HCT116 p53 −/− cells compared with HCT116 wild-type cells. Bars represent the mean±S.E.M. ( f ) The induction of p21 in Pttg2-treated cells occurs in a p53-dependent manner. Western blot analysis of p21 levels in HCT116 p53 −/− cells 72 h post-infection with shPttg2 or control lentivirus. β -Actin was used as loading control. ( g ) Role of p21 in anoikis in Pttg2-depleted cells. The percentage of apoptotic cells in HCT116 p21 −/− following treatment with shPttg2 was significantly reduced compared with HCT116 wild-type cells. Bars represent the mean±S.E.M.

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Inhibition, Infection, Western Blot, Control

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Loss of cell adhesion in shPttg2-treated cells precedes the onset of apoptosis. ( a ) Spheroid formation of Pttg2-depleted cells after z-VAD treatment. Wild-type HCT116 cells were infected for 72 h in the presence or absence of the caspases inhibitor z-VAD and the cell–cell adhesion capacity assayed on poly-HEMA (PH)-coated plates. The experiments were repeated three times, and representative areas of each culture are shown. ( b ) Cell-cycle profile of Pttg2-silenced HCT116 cells growing under poly-HEMA conditions after z-VAD treatment. The percentage of apoptotic cells (subG1) was significantly reduced in the presence of z-VAD, whereas the other cell-cycle phases (G1, S and G2/M) were not modified. Data are representative of two independent experiments. ( c ) Phase-contrast images to assess multicellular aggregates formation in shPttg2-treated HCT116 p53 −/− cells growing on poly-HEMA-coated plates. Images show a representative region of each condition. ( d ) Analysis of the cell-cycle profile of Pttg2-depleted HCT116 p53 −/− cells growing under poly-HEMA conditions. ShPttg2-treated cells lacking p53 showed a reduced percentage of cell death (subG1). The other cell-cycle phases (G1, S and G2/M) remained unaltered. Data are representative of two independent experiments

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Infection, Modification

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Pttg2-depleted cells exhibit cytoskeletal defects. Microtubule cytoskeleton of PTTG2 depleted cells. Confocal images of Pttg2-silenced HCT116 cells stably expressing Histone H2B (H2B)-cherry fusion gene. Seventy-two hours post-infection, cells were fixed with methanol at −20 °C and stained for α -tubulin (green). Scale bars, 25 μ m

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Stable Transfection, Expressing, Infection, Staining

Journal: Cell Death & Disease

Article Title: PTTG2 silencing results in induction of epithelial-to-mesenchymal transition and apoptosis

doi: 10.1038/cddis.2013.48

Figure Lengend Snippet: Pttg2 depletion results in deregulated E-cadherin and vimentin levels and induction of the epithelial-to-mesenchymal transition. ( a ) Summary of gene expression profile according to Gene Ontology. Pie charts show the percentage of genes included in each category (molecular function, biological process and cellular component) and subcategories with a P -value <0.01. The GO analysis was performed using DAVID v.6.7 program. GO analysis of differentially upregulated or downregulated genes are shown in upper and lower panels, respectively. ( b ) Validation of E-Cadherin downregulation in Pttg2-silenced cells by quantitative PCR. HCT116 cells were infected with shPttg2 or control lentivirus and assayed for E-cadherin mRNA levels 72 h post-infection. Values were normalized using the hprt1 gene as an internal control. Columns represent the mean values of 2–3 independent experiments ±S.E.M. ( c ) Western blot analysis of E-Cadherin repression in Pttg2-depleted HCT116 cells. The expression levels of the E-Cadherin protein in shPttg2 cells was compared with control cells expressing an empty lentiviral plasmid. Cells were grown either as a monolayer (attached) or in suspension for 48 h on poly-HEMA-coated plates (+PH) and total lysates analyzed by immunoblotting. ( d ) Vimentin mRNA induction in shPttg2-treated cells. HCT116 cells were infected with shPttg2 or control lentivirus and assayed for vimentin levels 72 h post-infection by quantitative PCR. Values were normalized using the hprt1 gene as an internal control. Columns represent the mean values of 2–3 independent experiments±S.E.M.

Article Snippet: Expression of Pttg2 and Pttg3 was measured using the Applied BiosystemsTaqMan Gene Expression Assays Hs00747713_sH and Hs00607485_s1.

Techniques: Gene Expression, Biomarker Discovery, Real-time Polymerase Chain Reaction, Infection, Control, Western Blot, Expressing, Plasmid Preparation, Suspension

Journal: PLoS ONE

Article Title: 5-ALA Fluorescence in Native Pituitary Adenoma Cell Lines: Resection Control and Basis for Photodynamic Therapy (PDT)?

doi: 10.1371/journal.pone.0161364

Figure Lengend Snippet: (A) The pituitary adenoma cell lines GH3 and AtT-20 were incubated for 6 and 24hrs with 100 g/ml 5-ALA (black line) or with media alone (gray area). One out of 3–7 independent experiments is shown. (B andC) MFI of all flow cytometry experiments are shown. Error bars represent S.E.M.

Article Snippet: The pituitary adenoma cell lines GH3 and AtT-20 (both from ATCC, Manassas, Virginia, US) and the glioblastoma cell line U373 MG (from the original cultures established in Uppsala 1968 [ ]) were cultured as adherent monolayer at 37àC in a 5% CO2 humidified atmosphere.

Techniques: Incubation, Flow Cytometry

Journal: PLoS ONE

Article Title: 5-ALA Fluorescence in Native Pituitary Adenoma Cell Lines: Resection Control and Basis for Photodynamic Therapy (PDT)?

doi: 10.1371/journal.pone.0161364

Figure Lengend Snippet: Comparison of flow cytometry, spectrofluorometer data and cell size. Comparison of flow cytometry, spectrofluorometer data and cell size in GH3, AtT-20 and U373MG cell lines after 6 and 24hrs of 5-ALA incubation. FACS as percent positive events n = 3–7. MFI as mean fluorescence intensity n = 3–7. Spec. as arbitrary units n = 3–4. Cell size n = 10

Article Snippet: The pituitary adenoma cell lines GH3 and AtT-20 (both from ATCC, Manassas, Virginia, US) and the glioblastoma cell line U373 MG (from the original cultures established in Uppsala 1968 [ ]) were cultured as adherent monolayer at 37àC in a 5% CO2 humidified atmosphere.

Techniques: Comparison, Flow Cytometry, Incubation, Fluorescence, FACS

Journal: PLoS ONE

Article Title: Magmas Overexpression Inhibits Staurosporine Induced Apoptosis in Rat Pituitary Adenoma Cell Lines

doi: 10.1371/journal.pone.0075194

Figure Lengend Snippet: (A) Expression of rat Magmas gene in GH1, GH3, GH4C1 and MMQ cell lines was determined by RT-qPCR and normalized to 18S rRNA as reference gene. Data are presented as fold change of mRNA levels of target gene (mean ± SE) in the cell lines vs. normal pituitary pool (NP), as detailed in Materials and Methods. *P < 0.05. (B) Expression of rat Magmas protein in GH3, GH4C1 and MMQ cell lines was determined by Western blot analysis and normalized to tubulin as housekeeping gene.

Article Snippet: GH4C1, GH3, GH1, and MMQ rat pituitary adenoma cell lines were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) repressed the growth and hormone secretion of pituitary adenomas. (a, b) GH3 and MMQ cells were treated with a range of concentrations of BT for 24 and 48 h. Cell viability was determined by MTS. (c, d) GH3 and MMQ cells were treated with BT for 24 h; formed colonies were photographed and counted using the ImageJ software. (e) Different subtypes of primary pituitary tumor cells were treated with 0.25 μ M BT for 24 h; cell viability was determined by MTS. (f) After treating GH3 and MMQ cells with BT (0–500 nM) for 24 h, culture medium supernatants were collected for ELISA to determine the prolactin hormone concentration. (g) After treating GH3 cells with BT (0–500 nM) for 24 h, culture medium supernatants were collected for ELISA to determine the levels of GH hormone. Data are represented as the mean ± SD. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Software, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) inhibits pituitary tumor growth in xenograft models. (a, d) Representative images of xenograft tumors from mice treated with control vehicle or BT at 24 days of drug administration. (b, e) Tumor volume growth curves of nude mice in different treatment groups. BT inhibited the rate of tumor growth in GH3 and MMQ cells. (c, f) Tumor weights from mice injected with GH3 cells and treated with control vehicle or BT at day 24 of drug administration. Data are represented as the mean ± SD. ∗ P < 0.05.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Control, Injection

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol- (BT-) induced apoptotic cell death in GH3 and MMQ cells. (a, b) Induction of apoptosis in GH3 and MMQ cells with BT treatment (250 nM) for 24 h followed by Annexin V and PI staining. Apoptosis ratios were measured by flow cytometry. (c, d) After 0–24 h treatment with BT (250 nM) in GH3 and MMQ cells, Western blot analysis was used to monitor the expression level of c-caspase-3, c-caspase-8, and Bcl-2. (e, f) GH3 and MMQ cells were treated with BT or NAC (100 μ M) alone or in combination for 24 h and MTS assays conducted to determine cell viability. Data are represented as the mean ± SD. ∗ P < 0.05.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Staining, Flow Cytometry, Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) downregulated the phosphorylation level of 4EBP1 and S6K1 in pituitary adenomas. (a) After 0–24 h treatment with BT (250 nM) in GH3 and MMQ cells, Western blot analysis was used to monitor the expression level of 4EBP1, S6K1, p-4EBP1, and p-S6K1. (b) Primary pituitary tumor cells (1 GHoma and 1 nonfunctional tumor) were treated with BT; Western blot analysis revealed the expression level of 4EBP1, S6K1, p-4EBP1, and p-S6K1. (c) Representative images of IHC-stained samples show that BT decreased p-4EBP1 and p-S6k1 expression in GH3 and MMQ xenograft models; scale bar, 100 μ m.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Phospho-proteomics, Western Blot, Expressing, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) increased ROS accumulation in GH3 and MMQ cells by inhibiting the Nrf2 pathway. (a) GH3 and MMQ cells treated with BT (250 nM) for 24 h were analyzed by flow cytometry after DCFH-DA staining to determine ROS levels. (b) GH3 and MMQ cells were treated with a range of concentrations of BT for 24 h. Western blot analysis showed that BT downregulated the expression of Nrf2 and its downstream gene Ho-1 but had no effect on Keap1. (c) GH3 and MMQ cell lines were treated with BT (250 nM) and/or NAC (100 μ M) for 24 h and cell viability measured by MTS assay. (d) GH3 and MMQ cell lines were treated with BT (250 nM) and/or NAC (100 μ M) for 24 h; Western blot analysis showed that NAC reversed the inhibitory effect of BT on inhibition of p-4EBP1 and p-S6K1. Data are presented as the mean ± SD. ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Flow Cytometry, Staining, Western Blot, Expressing, MTS Assay, Inhibition

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) increased the cytotoxicity of cabergoline in GH3 and MMQ cells. (a) GH3 and MMQ cells were treated with BT (250 nM) and/or cabergoline (50 μ M) for 24 h. Cell viability was measured by MTS assays, and (c) Western blot analysis was conducted to determine the level of p-4EBP1 and p-S6K1. (b) Representative images of colony formation assays. GH3 and MMQ cells were treated with BT (250 nM) and/or cabergoline (50 μ M) for 24 h. Colony formation was allowed to proceed for 14 days. Data are presented as the mean ± SD. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Brusatol Inhibits Tumor Growth and Increases the Efficacy of Cabergoline against Pituitary Adenomas

doi: 10.1155/2021/6696015

Figure Lengend Snippet: Brusatol (BT) augmented the cytotoxic effect of cabergoline in GH3 xenografts. (a) The 4 groups were treated with BT or/and cabergoline every other day. After 30 days of treatment, the nude mice were anesthetized, and the tumors were harvested. (b) Tumor volume was measured every 3 days (volume = π /6 × [larger diameter] × [smaller diameter] 2 ), and the volume curve was determined using GraphPad Prism 8.0. (c) Comparison of the average xenograft tumor weight. Data are represented as the mean ± SD. ∗ P < 0.05; ∗∗∗ P < 0.001.

Article Snippet: Rat pituitary tumor GH3 cell lines (ATCC CRL-10609) and MMQ cell lines (ATCC CRL-10609) were purchased from the American Type Culture Collection (ATCC).

Techniques: Comparison

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 1 The mRNA and protein expression of PTTG1 in pan-cancer. (A) The mRNA expression level of PTTG1 in tumor tissues and adjacent normal tissues in 33 TCGA dataset. The red or purple plots represent the tumor sample and blue plots represent normal sample, respectively. (B) The mRNA expression level of PTTG1 in tumor and normal tissues from TCGA and GTEx data sets. The red letters represent significantly high expression in tumor and green let ters represent low expression, respectively. (C) Pan-cancer analysis of PTTG1 gene expression at different tumor stages. (D) Immunohistochemical staining images of PTTG1 in human cancers and normal tissue from HPA database. *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing, Gene Expression, Immunohistochemical staining, Staining

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 2 Survival map of PTTG1 in pan-cancer. GEPIA2 survival analysis including OS. (A) and DFS (B) demonstrated the survival map (upper panel) and Kaplan–Meier survival plots (lower panel) of PTTG1 in TCGA dataset

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques:

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 3 Genetic alteration analysis of PTTG1 in various TCGA tumors. (A) The profile of SNV (Single Nucleotide Variation) of PTTG1 in the TCGA cohorts. (B) The lollipop plot presents the mutation site, type, and count of PTTG1 in the sample set of TCGA UCEC and STAD cohorts. (C) The SNV classes of PTTG1 in the TCGA cohorts. (Variant Classification): the count of each type of deleterious mutation; (Variant Type): the count of SNP and DEL of PTTG1; (SNV class): the count of each SNV class of PTTG1. (D) Pie plot summarizes the CNV (Copy Number Variation) of PTTG1 in the TCGA datasets. (E) The difference of survival between PTTG1 CNV and wide type in the TCGA datasets. (F) Overall survival (OS), progression free survival (PFS), disease free interval (DFI) and disease specific survival (DSS) of PTTG1 CNV and wide type in the TCGA KIRP cohort

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Mutagenesis, Variant Assay

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 4 PPI, GO and KEGG analysis of PTTG1 and similar genes. (A) Correlation analysis of PTTG1 and top 5 similar genes in TCGA database. (B) PPI network of PTTG1 and top 100 similar genes, where the MCODE complexes are colored according to their identities. (C) The five MCODE complexes identified in PPI network by Metascape, colored by their identities. The top-three functional enriched terms of each MOCDE network were listed at the right bottom. (D) Cellular component, molecular function, biological process, and KEGG analysis of PTTG1 and similar genes

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Functional Assay

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 5 Correlation analysis between PTTG1 expression and immune infiltration. (A) Heatmap summarizes the significance of P value and FDR for the spearman correlation analysis between PTTG1 expression and immune cells’ infiltrates (*: P value < 0.05; #: FDR ≤ 0.05). (B) The correlation between PTTG1 expression and ImmunoScore, StromalScore or ESTIMATE Score

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 6 Correlations between PTTG1 expression and immunotherapy response. (A) The plot showed the prognostic value of PTTG1 versus published biomarkers in melanoma and kidney renal clear cell carcinoma cohort. The x-axis shows the z-score on Cox-PH regression and the y-axis indicates its significance level (two-sided Wald test) (B) The association of PTTG1 with patients’ overall survival through Kaplan-Meier curves in melanoma and kidney renal clear cell carcinoma patients with immunotherapy treatment. (C) The bar chart showed the correlation between PTTG1 and published biomarkers in the immunotherapy cohorts. The AUC was used to evaluate the predictive performance of the test biomarker on the immunotherapy response state

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing, Biomarker Discovery

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 7 Mechanisms of PTTG1 promoting immunotherapy resistance. (A) The relationship between PTTG1 and CTL dysfunction. (B) Association of PTTG1 with immune checkpoint genes in pan-cancer. (C) The correlations between PTTG1 expression and TMB scores with Pearson correlation. (D) The correla tions between PTTG1 expression and HRD scores with Pearson correlation. *P < 0.05

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 8 Single-cell data analysis of PTTG1 expression by TISCH2. Cellular clusters and single-cell PTTG1 expression profile in different cellular types of NSCLC (A), KIRC (B), and LIHC (C). (D) Single-cell PTTG1 expression profile of non-responder and responder in LIHC patients with PD-L1/CTLA-4 treatment

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 9 PTTG1 increased the drug resistance of cancer cells. (A) The correlation between PTTG1 expression and the sensitivity [-log10(IC50)] of GDSC drugs. A depiction of the correlation between PTTG1 expression and MEK1/MEK2 inhibitor. (B) NSC-207895 (C) and ABT-263 (E) response obtained from the GDSC panel. (D) The predicted interaction of NSC-207895 with PTTG1 protein. Left: NSC-207895 binding with the pocket of PTTG1 is composed of hy drogen bonds. Right: the 2D hydrogen bond interaction pattern of NSC-207895 upon binding to PTTG1 protein. (E) The predicted interaction of ABT-263 with PTTG1 protein. Left: ABT-263 binding with the pocket of PTTG1 is composed of hydrogen bonds. Right: the 2D hydrogen bond interaction pattern of ABT-263 upon binding to PTTG1 protein

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Expressing, Binding Assay

Journal: BMC cancer

Article Title: Multi-omics analysis identifies PTTG1 as a prognostic biomarker associated with immunotherapy and chemotherapy resistance.

doi: 10.1186/s12885-024-13060-5

Figure Lengend Snippet: Fig. 10 The validation of PTTG1 characterization in lung adenocarcinoma. (A) The expression of PTTG1 was analyzed using the Wilcoxon signed-rank test in GSE32863, GSE19188 and GSE31210 NSCLC datasets. (B) Kaplan–Meier curves and log-rank test of PTTG1 in NSCLC data sets. (C) The mRNA expres sion of PTTG1 in H1299, H1355 parental and resistance cells in GSE77209 dataset. T[n]: Resistant cells generated after ‘n’ cycles of paclitaxel-carboplatin treatment. (D) The mRNA expression levels of PTTG1 in parental cells A549, H358 and paclitaxel resistant cells A549- TXR, H358- TXR were detected by RT‑qPCR. (E) PTTG1 siRNA knockdown efficiency in A549- TXR and H358- TXR measured. (F) PTTG1, CDC2 and CCNB1 were upregulated in A549- TXR and H358- TXR. (G) Clone formation assays with A549- TXR and H358- TXR cells transfected with PTTG1 siRNA. (H) Transwell assays with A549- TXR and H358- TXR cells transfected with PTTG1 siRNA. (I) PTTG1, CDC2 and CCNB1 were decreased after PTTG1 siRNA treatment measured by Western blot in A549-TXR and H358-TXR cells. *P < 0.05, **P < 0.01

Article Snippet: After blocking the membranes with 5% nonfat milk, incubating membranes with primary antibodies against Cyclin B1 (1:1000, Cat. 4138, Cell signaling), CDC2 (1:1000, Cat. 9116, Cell signaling), PTTG1 (1:1000, Cat. 18040-1-AP, Proteintech), GAPDH (1:1000, Cat. 2118, Cell signaling) overnight at 4 °C.

Techniques: Biomarker Discovery, Expressing, Generated, Knockdown, Transfection, Western Blot