Journal: Viruses

Article Title: Celecoxib Inhibits the Lytic Activation of Kaposi’s Sarcoma-Associated Herpesvirus through Down-Regulation of RTA Expression by Inhibiting the Activation of p38 MAPK

doi: 10.3390/v7052268

Figure Lengend Snippet: COX-2 inhibitors, NS-398 and nimesulide, have no effect on KSHV lytic replication in BCBL-1 cells. The cells induced with TPA for 3 h were treated with the test compounds for 72 h. Then, the effect of NS-398 and nimesulide on cell viability ( A , C ) and the number of DNA copies ( B , D ) were measured as described in the Methods. CDV (20 µM) is a positive control for inhibiting viral DNA replication. Data were normalized as the fold change compared with the no-TPA induced control. The results are presented as the mean values with standard deviations ( n = 3).

Article Snippet: Celecoxib was purchased from Sigma-Aldrich (St. Louis, MO, USA), NS-398, SP600125 and SB203580 were purchased from Beyotime Institute of Biotechnology, and nimesulide and cidofovir (CDV) were obtained from Selleck Chemicals (Shanghai, China).

Techniques: Positive Control, Control

Journal: Viruses

Article Title: Celecoxib Inhibits the Lytic Activation of Kaposi’s Sarcoma-Associated Herpesvirus through Down-Regulation of RTA Expression by Inhibiting the Activation of p38 MAPK

doi: 10.3390/v7052268

Figure Lengend Snippet: COX-2 inhibitors, NS-398 and Nimesulide, have no effect on KSHV reactivation in iSLK.219 cells. The cells were lytically induced with Dox and NaB in the presence of various concentrations of test compounds and incubated for 24 h. The effect of NS-398 and nimesulide on cytotoxicity to cells ( A , C ) and the fluorescence intensity of RFP ( B , D ) were detected and digitalized as described in the Methods. SP600125 (5 µg/mL) is a positive control for inhibiting KSHV reactivation. Data were normalized as the relative fluorescence intensity compared to the DMSO control. The results are presented as the mean values with standard deviations ( n = 3).

Article Snippet: Celecoxib was purchased from Sigma-Aldrich (St. Louis, MO, USA), NS-398, SP600125 and SB203580 were purchased from Beyotime Institute of Biotechnology, and nimesulide and cidofovir (CDV) were obtained from Selleck Chemicals (Shanghai, China).

Techniques: Incubation, Fluorescence, Positive Control, Control

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 1 Upregulation of GRP78 by NSAIDs. AGS (a and c) or MKN-45 and Kato III (b) cells were incubated with indicated concentrations of stated NSAIDs for 12 h (celecoxib) or 24 h (NSAIDs other than celecoxib). Cells were pretreated with indicated concentrations of PGE2 for 2 h before the celecoxib treatment (c). Whole cell extracts (5 mg protein) were analysed by immunoblotting with an antibody against GRP78 or actin. Band intensity of GRP78 was determined by densitometric scanning, gel- loading levels compensated against the band intensity of actin, and expressed relative to the control sample (i.e. without NSAIDs).

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Incubation, Western Blot, Control

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 2 Upregulation of ER chaperone genes by celecoxib. AGS cells were incubated with indicated concentrations (a) or 100 mM (b) of celecoxib for 12 h (a) or the time periods indicated (b) and total RNA extracted. Samples were subjected to real-time RT–PCR by use of a specific primer for each gene. Values were normalized to actin gene expression and expressed relative to the control sample (i.e. without celecoxib). Values given are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Incubation, Quantitative RT-PCR, Gene Expression, Control

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 3 Effect of celecoxib on growth of xenograft tumor and expression of GRP78 in nude mice. Each nude mouse was inoculated s.c. with MKN-45 cells and tumors were developed until size of tumors reached a mean volume of 116734 mm3. Then indicated dose of celecoxib was administered single daily orally for the duration of the study. Tumors were measured weekly and their volumes calculated (a). After 4 days from the start of celecoxib administration, cell lyzates prepared from tumors were analysed by immunoblotting with an antibody against GRP78 or actin (b). Band intensity of GRP78 was determined by densitometric scanning, compensated against the band intensity of actin, and expressed relative to the control sample (i.e. without celecoxib) (c). Values given are mean7s.d. (n ¼ 6 for (a) and n ¼ 3 for (b and c)). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Expressing, Western Blot, Control

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 4 Activation of PERK, eIF2a and ATF4 by celecoxib. AGS cells were incubated with 100 mM (a, c and e) or indicated concentrations (b and d) of celecoxib for the time periods indicated (a, c and e) or 12 h (b and d). For (a, d and e), whole-cell extracts (5 mg protein for actin, 10 mg protein for ATF4 and 20 mg protein for PERK and eIF2a) were analysed by immunoblotting with an antibody against phosphorylated PERK (p-PERK), phosphory- lated eIF2a (p-eIF2a), ATF4 or actin. For (b and c), total RNA was extracted and subjected to real-time RT–PCR by use of a specific primer for ATF4. Values were analysed and expressed as previously described in the legend of Figure 2. Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Activation Assay, Incubation, Western Blot, Quantitative RT-PCR

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 5 Effect of siRNA for ATF4 on the celecoxib-dependent upregulation of GRP78. AGS cells transfected with siRNA for ATF4 (siATF4) or nonsilencing (ns) siRNA were incubated with or without 100 mM celecoxib for 12 h. Total RNA was extracted and subjected to real-time RT–PCR by use of a specific primer for ATF4 (a), GRP78 (b), ATF2 (c) and ATF3 (d). Values were analysed and expressed as previously described in the legend of Figure 2. Values shown are mean7s.d. (n ¼ 3). ***Po0.001; *Po0.05. n.s., not significant.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Transfection, Incubation, Quantitative RT-PCR

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 6 Changes in intracellular Ca2 þ concentration in the NSAID-dependent upregulation of GRP78. The intracellular Ca2 þ concentration was monitored by a fluo-3/AM assay system. Indicated concentrations of celecoxib were added to fluo-3/AM- loaded cells and the time–course of fluo-3 fluorescence change monitored. The maximum value for the increase in the intracellular Ca2 þ level (D[Ca2 þ]i) is shown (a). AGS cells were preincubated with or without 2 mM BAPTA-AM for 1 h and further incubated with or without 80 mM celecoxib (b–d), 800 mM nimesulide (e), 800 mM diclofenac (f) or 400 mM indomethacin (g) in the presence or absence of 2 mM BAPTA-AM for 6 h (celecoxib) or 12 h (other NSAIDs). The levels of GRP78 protein (b, e–g), GRP78mRNA (c) and ATF4 mRNA (d) were estimated by immunoblotting or real- time RT–PCR experiments as described in the legends of Figures 1 and 2. AGS cells were incubated with 100 mM celecoxib or 2 mM thapsigargin for indicated periods (h). Whole cell extracts (25 mg protein for ATF6 and 10 mg protein for actin) (upper panel in (h)) or nuclear extracts (20 mg protein for p50 ATF6 and 5 mg protein for lamin B) (lower panel in (h)) were analysed by immunoblotting with an antibody against ATF6, actin or lamin B as described in the legends of Figure 1. As for p50 ATF6 band, intensity of each band was expressed relative to the positive control sample (i.e. cells treated with thapsigargin for 1.5 h). Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Concentration Assay, Incubation, Western Blot, Quantitative RT-PCR, Positive Control

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 7 Induction of apoptosis by celecoxib. AGS cells were incubated with the indicated concentrations (a and c) or 100 mM (b and d) of celecoxib for 12 h (a and c) or indicated periods (b and d). For (a and b), apoptotic cell numbers were determined by FACS (a and b). For (c and d) total RNA was extracted and subjected to real-time RT–PCR by use of a specific primer for CHOP. Values were analysed and expressed as previously described in the legend of (Figure 2c and d). Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Incubation, Quantitative RT-PCR

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 8 Effect of overexpression of GRP78 on celecoxib-induced apoptosis. AGS cells were transfected with the indicated amount (a and b) or 0.5 mg (c and d) of plasmid for the overexpression of GRP78 and pcDNA3.1 vector (total DNA amounts were fixed at 4 mg). After 48 h, cells were incubated with or without100 mM celecoxib for 6 h (b–d). The levels of GRP78 protein (a), GRP78mRNA (c) and CHOP mRNA (d) were estimated by immunoblotting or real-time RT–PCR experiments as previously described in the legends of Figures 1 and 2. Apoptotic cell numbers were determined by FACS as described in the legend of Figure 7(b). Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Over Expression, Transfection, Plasmid Preparation, Incubation, Western Blot, Quantitative RT-PCR

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 9 Stimulation of the antiapoptotic effect of GRP78 against celecoxib by coexpression of ERdj3 or ERdj4. AGS cells were transfected with the indicated amounts of each expression plasmid and pcDNA3.1 vector (total DNA amounts were fixed at 4 mg). After 48 h, AGS cells were incubated with or without 100 mM celecoxib for 6 h. Apoptotic cell numbers were determined by FACS as described in the legend of Figure 7. Values shown are mean7s.d. (n ¼ 3). ***, www and ###Po0.001; **, ww and ##Po0.01; * and #o0.05. * (versus celecoxib-treated cells only), # (versus celecoxib-treated and GRP78 overexpressing cells), w (versus celecoxib-treated and ERdj3 (or ERdj4) overexpressing cells). n.s, not significant (a–d).

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Transfection, Expressing, Plasmid Preparation, Incubation

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 10 Effect of GRP78 siRNA on celecoxib-induced apopto- sis. AGS cells were transfected with 5 mg of siRNA for GRP78 (siGRP78) or nonsilencing (ns) siRNA. After 48 h, cells were incubated with or without 80 mM celecoxib (a–d and f), indicated concentrations of celecoxib (e) or indicated concentrations of staurosporine (g and h) for 6 h. The levels of GRP78 protein (a and h), GRP78 mRNA (b), ERdj3 mRNA (c), ERdj4 mRNA (d) and CHOP mRNA (f) were estimated by immunoblotting or real-time RT–PCR experiments as described in the legends of Figures 1 and 2. Apoptotic cell numbers were determined by FACS as described in the legend of (Figure 7e and g). Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Transfection, Incubation, Western Blot, Quantitative RT-PCR

Journal: Science translational medicine

Article Title: Advances toward transformative therapies for tendon diseases

doi: 10.1126/scitranslmed.abl8814

Figure Lengend Snippet: Current therapies in clinical use for tendon diseases. Examples of products of a given therapeutic modality and mechanism of action are provided. GCR, Glucocorticoid Receptor.

Article Snippet: Therapeutic modality Molecular/biological mechanism of action Product Company Launch Chemical drug NSAID/anti-inflammatory, analgesic Indocin (indomethacin) GSK 1965 Mesulid (nimesulide) Boehringer Ingelheim, GSK, Merck, Pfizer 1985 Aulin (nimesulide) Sanofi 1990 Duraprox (oxaprozin) Aventis 2002 Felbinac (felbinac) Toko Pharmaceutical Industrial 2018 Chemical drug Corticosteroid/anti-inflammatory, analgesic Kenalog (triamcinolone acetonide) Apothecon 1978 Chemical drug Cox-2 inhibitor/anti-inflammatory, analgesic Celebrex (celecoxib) Astella Pharma 2009 Chemical drug GCR agonist/anti-inflammatory, analgesic Lederlon (triamcinolone hexacetonide) Esteve 2001 Lederspan (triamcinolone hexacetonide) Mylan Biological product PRP Not specified/tissue regeneration Terumo BCT (PRP) Terumo 2010 Cell therapy Not specified/tissue regeneration Ortho-ATI (autologous tendon cells) Johnson & Johnson, Orthocell 2010 Biomaterial Not specified/viscosupplement CartiZol (atelocollagen) Sewon Cellontech 2007 Ostenil (hyaluronan) TRB Chemedica 2012 OrthoVisc (hyaluronan) Anika Therapeutics 2016 Open in a separate window Current therapies in clinical use for tendon diseases.

Techniques: