Journal: Neural Regeneration Research

Article Title: Subretinal fibrosis secondary to neovascular age-related macular degeneration: mechanisms and potential therapeutic targets

doi: 10.4103/NRR.NRR-D-23-01642

Figure Lengend Snippet: Molecules and pathways promoting subretinal fibrosis, as well as the potential drugs, are depicted. Created with BioRender.com. COX2: Cyclooxygenase-2; IL-6: interleukin 6; IL-6R: interleukin-6 receptor; MRTF-A: myocardin-related transcription factor A; PAF-R: platelet-activating factor receptor; PDGF: platelet-derived growth factor; PDGFR-β: platelet-derived growth factor receptor-β; (P)RR: (pro)renin receptor; (P)RR-PshRNA: (P)RR-proline-modified short hairpin RNA; RAR: retinoic acid receptor; siRNA: small interfering RNA; SRF: serum response factor; TGFβ: transforming growth factor-β; YAP: Yes-associated protein.

Article Snippet: Pro-fibrotic , Transforming growth factor-β signaling pathway Wnt signaling pathway Phosphatidylinositol-3-kinase/Akt pathway Vascular endothelial growth factor/vascular endothelial growth factor receptor Platelet derived growth factor/platelet derived growth factor receptor-β Interleukin-6/interleukin-6 receptor Hypoxia-inducible factor-1α/p53/miRNA-34a/Klotho pathway MRTF-A-SRF pathway Connective tissue growth factor, fibroblast growth factor 2, platelet-activating factor receptor, Galectin-1, Yes-associated protein, adiponectin, methyltransferase-like 3, matrix metalloproteinase 12, peptidyl arginine deiminase-4, αB-Crystallin, Snail (SNIA1), cyclooxygenase-2, (pro)renin receptor, sphingosine-1-phosphate, γ-secretase.

Techniques: Derivative Assay, Modification, shRNA, Small Interfering RNA

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: PTTG1 expression in OSCC tissues and cells. (A) Representative images (left) and quantification (right) of PTTG1 immunohistochemistry staining in healthy (n=32) and OSCC tissues (n=32). Scale bar, 100 µm; original magnification, ×20. (B) PTTG1 expression was analyzed by RT-qPCR for the pooled OSCC tissue samples. *P<0.05, **P<0.01 and ***P<0.001 vs. non-tumor. (C) Protein expression of PTTG1 in the pooled OSCC samples revealed by western blotting. The upper panel shows the membranes stained with antibodies, with GAPDH as an internal control, and the lower panel shows the relative quantification of protein expression. (D) mRNA and (E) protein expression (left) and fold change (right) of PTTG1 and p21 were analyzed by RT-qPCR and western blotting in OSCC cell lines. *P<0.05, **P<0.01 and ***P<0.001 vs. HSC-2 cell lines using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; RT-qPCR, reverse transcription quantitative PCR; N, non-tumor; T, tumor.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Immunohistochemistry, Staining, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: PTTG1 expression in relation to cell proliferation, cell cycle and apoptosis in OSCC cells. (A) PTTG1 expression was analyzed by reverse transcription quantitative PCR in OSCC cells. (B) Protein expression of PTTG1 and PCNA in OSCC cell lines revealed by western blotting. The left panel shows the membranes stained with antibodies, with GAPDH as an internal control, and the right panel shows the relative quantification of protein expression. (C and D) Representative images of cell proliferation ability (left) and quantification (right) of PTTG1 using the EdU assay in the (C) HSC-2 and (D) SCC-9 cell lines (scale bars, 100 µm and 50 µm, respectively; original magnification, ×40). The percentages of HSC-2 and SCC-9 cells treated with VC or siR-PTTG1 were determined by EdU incorporation (green) and DAPI (blue). (E) Protein expression (left) and the fold change (right) of cell cycle markers, including cyclins D1, E, and B1 in OSCC cells. (F) Protein expression (left) and fold change (right) related to apoptosis markers, including Cas-7, c-Cas-7, and c-PARP in OSCC cells. *P<0.05, **P<0.01 and ***P<0.001 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; PCNA, proliferating cell nuclear antigen; VC, vehicle control; siR-PTTG1, small interfering RNA-PTTG1; EdU, 5-ethynyl-2′-deoxyuridine; Cas-7, Caspase-7; c-, cleaved-; PARP, poly (ADP-ribose) polymerase.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, EdU Assay, Small Interfering RNA

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: The effect of PTTG1 on cellular senescence and DNA damage in OSCC. (A) mRNA expression of p21 was analyzed by reverse transcription quantitative PCR in OSCC cells. (B) Protein expression (left) and quantification (right) of PTTG1 and p21 were analyzed by western blotting in OSCC cells. (C) Representative images (left) and numbers (right) of cellular senescence in OSCC cells detected by senescence-associated β-galactosidase staining (blue). The percentages of HSC-2 and SCC-9 were determined by β-galactosidase incorporation (scale bars, 50 µm and 200 µm, respectively; original magnification, ×40). (D) Protein expression (left) and fold change (right) related to DNA damage, including γH2AX, p-ATR and p-ATM in OSCC cells. GAPDH was used as an internal control. (E) Representative images (left) and numbers (right) of chromosomal damage detected by γH2AX staining in HSC-2 cells (scale bar, 50 µm; original magnification, ×63). (F) Representative images (left) and numbers (right) of chromosomal damage detected by γH2AX staining in SCC-9 cells (scale bar, 250 µm; original magnification, ×63). *P<0.05, **P<0.01 and ***P<0.001 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; γH2AX, phosphorylated histone H2AX; ATR, ataxia telangiectasia and Rad3-related protein; p-, phosphorylated; ATM, ataxia telangiectasia mutant; siR-PTTG1, small interfering RNA-PTTG1; SA-β gal, senescence-associated beta-galactosidase; VC, vehicle control.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, Mutagenesis, Small Interfering RNA

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: The effect of PTTG1 downregulation on tumor growth in vivo . (A) A schematic diagram of transfection in vivo . (B) Representative images of tumor sizes in vivo . The tumor size of each group was depicted graphically in vivo . (scale bar, 500 mm). (C) Tumor weights of each group in vivo . (D) Protein expression (left) and fold change (right) related to cell proliferation, apoptosis, cellular senescence and DNA damage, including PCNA, c-Cas-7, p21, γH2AX and p-ATM in OSCC cell lines. GAPDH was used as an internal control. (E and F) Representative images (left) and numbers (right) of apoptotic DNA damage analyzed by TUNEL assay (red) and DAPI (blue) in (E) HSC-2 and (F) SCC-9 cells (scale bar, 50 µm; original magnification, ×20). *P<0.05, **P<0.01, ***P<0.001, # P<0.05 and ## P<0.01 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; PCNA, proliferating cell nuclear antigen; c-Cas-7, cleaved Caspase-7; γH2AX, phosphorylated histone H2AX; ATM, ataxia telangiectasia mutant; p-, phosphorylated; Cas-7, Caspase-7; ATR, ataxia telangiectasia and Rad3-related protein; ATM, ataxia telangiectasia mutant; OSCC, oral squamous cell carcinoma; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; VC, vehicle control.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: In Vivo, Transfection, Expressing, Control, TUNEL Assay, Mutagenesis, End Labeling

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: A schematic model of DNA damage induced by the downregulation of PTTG1 in oral squamous cell carcinoma. PTTG1, pituitary tumor-transforming gene 1; phosphorylated histone H2AX; p-ATR, Phosphorylated ataxia telangiectasia and Rad3-related protein; p-ATM, phosphorylated ataxia telangiectasia mutant; SASP, senescence-associated secretory phenotype; c-Cas-7, cleaved Caspase-7; c-PARP, cleaved poly (ADP-ribose) polymerase.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Mutagenesis

Journal: JBMR Plus

Article Title: IL-33 prevents age-related bone loss and memory impairment by suppression of Th17 response: evidence in a d -galactose–induced aging mouse model

doi: 10.1093/jbmrpl/ziae101

Figure Lengend Snippet: IL-33 ameliorates d -galactose (D-gal)–induced suppression of osteoblast differentiation, represses osteoblast senescence, and attenuates inflammatory responses in D-gal–stimulated osteoblasts by modulating expression of IL-17. (A) Alkaline phosphatase (ALP) activity was measured at 72 hours after D-gal administration to assess cell differentiation. (B) An MTT 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide test was used to determine cell viability at 72 hours after D-gal treatment. (C) ALP activity in the presence of IL-33 with positive control BMP-2. Data are expressed as mean ± SEM ( n = 3). * p < .05, ** p < .01, and *** p < .005 compared with control. $ p < .05, $$ p < .01, and $$$ p < .005 compared with D-gal. # p < .05, ## p < .01, and ### p < .005 compared with D-gal + BMP-2. (D) Representative images of alizarin Red staining showing mineralization among different groups. (E) Quantification of mineralization using 10% CPC cetylpyridinium chloride. (F) Protein expression of Runx-2, type 1 collagen, P53, P21, pRB, PCNA, and Ki-67 gene in primary OB cells on treatment with D-gal and IL-33. (G–M) Quantification of protein expression using ImageJ software. (N–R) Relative mRNA expression of inflammatory cytokines such as IL-1β, TNF-α, and IL-17, IL-33, and IL-10 after 72 hours of treatment with D-gal, IL-17, and IL-33. Data expressed as mean ± SEM; n = 3. * p < .05, ** p < .01, and *** p < .005 compared with control. $ p < .05, $$ p < .01, and $$$ p < .005 compared with D-gal.

Article Snippet: We bought pRB Phospho-Rb, p53, p21, PCNA Proliferating cell nuclear antigen, Runx-2, and type 1 collagen from CST Cell Signaling Technology.

Techniques: Expressing, Activity Assay, Cell Differentiation, Positive Control, Control, Staining, Software

Journal: JBMR Plus

Article Title: IL-33 prevents age-related bone loss and memory impairment by suppression of Th17 response: evidence in a d -galactose–induced aging mouse model

doi: 10.1093/jbmrpl/ziae101

Figure Lengend Snippet: Effect of IL-33 on senescence and osteogenic and neurological markers. (A) Relative mRNA expression of senescence and osteogenic genes such as Runx-2, P53, and P21 in different groups. (B) Protein expression of Runx-2, type 1 collagen (T1Col), P53, P21, and pRB genes in different groups. (C–G) Quantification of protein expression using ImageJ software. (H) Serum beta-galactosidase (βGAL) levels. (I) Serum PTH levels. Data are expressed as mean ± SEM; n = 3. * p < .05, ** p < .01, and *** p < .005 compared with control. $ p < .05, $$ p < .01, and $$$ p < .005 compared with d -galactose (D-gal). (J) Illustrative immunoblots show expression of BACE1, p-tau, tau, p-CREB, CREB, and β-actin in cortex and hippocampus regions. The bar graphs show quantification of relative protein density of (K) BACE-1, (L) p-tau, and (M) p-CREB in the cortex and hippocampus regions after normalization with β-actin, t-tau, and t-CREB, respectively. (K–M) Data are expressed as mean ± SEM of n = 3 mice/group. Data were analyzed by repeated-measures 2-way ANOVA and 1-way ANOVA, followed by Bonferroni post hoc test. * P < .0332, ** p < .0021, *** p < .0002, **** p < .0001; $ p < .0332, $$ p < .0021, $$$ p < .0002, $$$$ p < .0001. * Control vs D-gal; D-gal vs IL-33.

Article Snippet: We bought pRB Phospho-Rb, p53, p21, PCNA Proliferating cell nuclear antigen, Runx-2, and type 1 collagen from CST Cell Signaling Technology.

Techniques: Expressing, Software, Control, Western Blot

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: PTTG1 expression in OSCC tissues and cells. (A) Representative images (left) and quantification (right) of PTTG1 immunohistochemistry staining in healthy (n=32) and OSCC tissues (n=32). Scale bar, 100 µm; original magnification, ×20. (B) PTTG1 expression was analyzed by RT-qPCR for the pooled OSCC tissue samples. *P<0.05, **P<0.01 and ***P<0.001 vs. non-tumor. (C) Protein expression of PTTG1 in the pooled OSCC samples revealed by western blotting. The upper panel shows the membranes stained with antibodies, with GAPDH as an internal control, and the lower panel shows the relative quantification of protein expression. (D) mRNA and (E) protein expression (left) and fold change (right) of PTTG1 and p21 were analyzed by RT-qPCR and western blotting in OSCC cell lines. *P<0.05, **P<0.01 and ***P<0.001 vs. HSC-2 cell lines using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; RT-qPCR, reverse transcription quantitative PCR; N, non-tumor; T, tumor.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Immunohistochemistry, Staining, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: PTTG1 expression in relation to cell proliferation, cell cycle and apoptosis in OSCC cells. (A) PTTG1 expression was analyzed by reverse transcription quantitative PCR in OSCC cells. (B) Protein expression of PTTG1 and PCNA in OSCC cell lines revealed by western blotting. The left panel shows the membranes stained with antibodies, with GAPDH as an internal control, and the right panel shows the relative quantification of protein expression. (C and D) Representative images of cell proliferation ability (left) and quantification (right) of PTTG1 using the EdU assay in the (C) HSC-2 and (D) SCC-9 cell lines (scale bars, 100 µm and 50 µm, respectively; original magnification, ×40). The percentages of HSC-2 and SCC-9 cells treated with VC or siR-PTTG1 were determined by EdU incorporation (green) and DAPI (blue). (E) Protein expression (left) and the fold change (right) of cell cycle markers, including cyclins D1, E, and B1 in OSCC cells. (F) Protein expression (left) and fold change (right) related to apoptosis markers, including Cas-7, c-Cas-7, and c-PARP in OSCC cells. *P<0.05, **P<0.01 and ***P<0.001 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; PCNA, proliferating cell nuclear antigen; VC, vehicle control; siR-PTTG1, small interfering RNA-PTTG1; EdU, 5-ethynyl-2′-deoxyuridine; Cas-7, Caspase-7; c-, cleaved-; PARP, poly (ADP-ribose) polymerase.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, EdU Assay, Small Interfering RNA

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: The effect of PTTG1 on cellular senescence and DNA damage in OSCC. (A) mRNA expression of p21 was analyzed by reverse transcription quantitative PCR in OSCC cells. (B) Protein expression (left) and quantification (right) of PTTG1 and p21 were analyzed by western blotting in OSCC cells. (C) Representative images (left) and numbers (right) of cellular senescence in OSCC cells detected by senescence-associated β-galactosidase staining (blue). The percentages of HSC-2 and SCC-9 were determined by β-galactosidase incorporation (scale bars, 50 µm and 200 µm, respectively; original magnification, ×40). (D) Protein expression (left) and fold change (right) related to DNA damage, including γH2AX, p-ATR and p-ATM in OSCC cells. GAPDH was used as an internal control. (E) Representative images (left) and numbers (right) of chromosomal damage detected by γH2AX staining in HSC-2 cells (scale bar, 50 µm; original magnification, ×63). (F) Representative images (left) and numbers (right) of chromosomal damage detected by γH2AX staining in SCC-9 cells (scale bar, 250 µm; original magnification, ×63). *P<0.05, **P<0.01 and ***P<0.001 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; OSCC, oral squamous cell carcinoma; γH2AX, phosphorylated histone H2AX; ATR, ataxia telangiectasia and Rad3-related protein; p-, phosphorylated; ATM, ataxia telangiectasia mutant; siR-PTTG1, small interfering RNA-PTTG1; SA-β gal, senescence-associated beta-galactosidase; VC, vehicle control.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Staining, Control, Mutagenesis, Small Interfering RNA

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: The effect of PTTG1 downregulation on tumor growth in vivo . (A) A schematic diagram of transfection in vivo . (B) Representative images of tumor sizes in vivo . The tumor size of each group was depicted graphically in vivo . (scale bar, 500 mm). (C) Tumor weights of each group in vivo . (D) Protein expression (left) and fold change (right) related to cell proliferation, apoptosis, cellular senescence and DNA damage, including PCNA, c-Cas-7, p21, γH2AX and p-ATM in OSCC cell lines. GAPDH was used as an internal control. (E and F) Representative images (left) and numbers (right) of apoptotic DNA damage analyzed by TUNEL assay (red) and DAPI (blue) in (E) HSC-2 and (F) SCC-9 cells (scale bar, 50 µm; original magnification, ×20). *P<0.05, **P<0.01, ***P<0.001, # P<0.05 and ## P<0.01 vs. VC using Student's t-test. All experiments were performed in triplicate. PTTG1, pituitary tumor-transforming gene 1; PCNA, proliferating cell nuclear antigen; c-Cas-7, cleaved Caspase-7; γH2AX, phosphorylated histone H2AX; ATM, ataxia telangiectasia mutant; p-, phosphorylated; Cas-7, Caspase-7; ATR, ataxia telangiectasia and Rad3-related protein; ATM, ataxia telangiectasia mutant; OSCC, oral squamous cell carcinoma; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; VC, vehicle control.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: In Vivo, Transfection, Expressing, Control, TUNEL Assay, Mutagenesis, End Labeling

Journal: Oncology Reports

Article Title: Pituitary tumor‑transforming gene 1 regulates the senescence and apoptosis of oral squamous cell carcinoma in a p21‑dependent DNA damage response manner

doi: 10.3892/or.2024.8794

Figure Lengend Snippet: A schematic model of DNA damage induced by the downregulation of PTTG1 in oral squamous cell carcinoma. PTTG1, pituitary tumor-transforming gene 1; phosphorylated histone H2AX; p-ATR, Phosphorylated ataxia telangiectasia and Rad3-related protein; p-ATM, phosphorylated ataxia telangiectasia mutant; SASP, senescence-associated secretory phenotype; c-Cas-7, cleaved Caspase-7; c-PARP, cleaved poly (ADP-ribose) polymerase.

Article Snippet: The p53 mutant human OSCC cell lines HSC-2, SCC-9 and YD-10B were obtained from the Japanese Collection of Research Bioresources Cell Bank, the American Type Culture Collection and the Korean Cell Line Bank, respectively.

Techniques: Mutagenesis