Journal: British Journal of Cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: (A) Pentraxin 3 production levels of supernatants used for culturing of pancreatic carcinoma cell lines. ( B , C ) Cell migration assay in the presence or absence of the indicated reagents. NT (non-treated), FBS, 20% in the lower chamber; VEGF, 50 ng ml −1 in the lower chamber; PTX3, 50–100 ng ml −1 ; CRP, 50–100 ng ml −1 . Statistical significance was evaluated by comparison with or without the presence of PTX3 and CRP.

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques: Cell Migration Assay

Journal: British Journal of Cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: (A) Pentraxin 3 production levels of supernatants used for culturing of PTX3 and NT-control transfectant of pancreatic carcinoma cell lines. ( B ) Cell migration assay using pancreatic carcinoma cell lines (AsPC-1 and PANC-1) transfected with pCMV6-entry PTX3 ORF in the presence or absence of the indicated reagents. NT (non-treated), FBS, 20% in the lower chamber; VEGF, 50 ng ml −1 in the lower chamber; PTX3, 50–100 ng ml −1 ; CRP, 50–100 ng ml −1 . Stable transformants were subjected to cell migration assay.

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques: Transfection, Cell Migration Assay

Journal: British Journal of Cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: Patient demographic and clinical characteristics

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques:

Journal: British Journal of Cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: Kaplan–Meier curves for (A) progression-free survival according to blood-PTX3 level and (B) overall survival according to blood-PTX3 level. Cutoff points for PTX3 level were based on mean PTX3 level.

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques:

Journal: British Journal of Cancer

Article Title: Clinical impact of pentraxin family expression on prognosis of pancreatic carcinoma

doi: 10.1038/bjc.2013.348

Figure Lengend Snippet: Results of univariate and multivariate analyses

Article Snippet: We used the transfectant pCMV6-entry PTX3 open reading frame (ORF) clones (OriGene Technologies, Inc., Rockville, MD, USA; cat. no. RC207922) to investigate the cellular activity induced by intracellular PTX3 overexpression.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: K284-6111 alleviates memory impairment and neuroinflammation in Tg2576 mice by inhibition of Chitinase-3-like 1 regulating ERK-dependent PTX3 pathway

doi: 10.1186/s12974-020-02022-w

Figure Lengend Snippet: Effect of K284-6111 on PTX3-involved neuroinflammation. a Gene network analysis associated with CHI3L1 was carried out using the web-based analysis tool. BV-2 cells were treated with Aβ (5 μM) and K284-6111 (2 μM). (B) The mRNA expression level of Chi3l1 and Ptx3 in BV-2 cells was assessed by qRT-PCR. Expression of PTX3 was detected by Western blot ( c ) in the Tg2576 mice brain and ( d ) in the BV-2 cells. BV-2 cells were transfected with PTX3 siRNA (20 nM). After 24 hr, cells were treated with Aβ (5 μM). e The mRNA expression level of pro-inflammatory cytokines ( Tnf , Il1b , and Il6 ) and M1 microglia phenotype marker ( Cd86 ) in BV-2 cells were assessed by qRT-PCR. f Expression of iNOS and COX-2 was detected by Western blot. (G) Level of p-ERK 1/2, ERK 1/2, p-IκBα, and IκBα were detected by Western blot. BV-2 cells were treated with Aβ (5 μM), K284-6111 (2 μM), U0126 (20 μM), and Bay 11-7082 (5 μM). (H) Expression of PTX3 were detected by Western blot. BV-2 cells were transfected with CHI3L1 plasmid vector. After 24 hr, cells were treated with Aβ (5 μM), K284-6111 (2 μM), U0126 (20 μM), and Bay 11-7082 (5 μM). (I) Expression of PTX3 were detected by Western blot.

Article Snippet: Negative control (NC), PTX3 siRNA were purchased from OriGene Technologies, Inc. (Rockville, MD, USA). pcDNA3.1(+)-6 × Myc-CHI3L1 vector was cloned from Bionics (Seoul, Republic of Korea).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Marker, Plasmid Preparation

Journal: Journal of Neuroinflammation

Article Title: K284-6111 alleviates memory impairment and neuroinflammation in Tg2576 mice by inhibition of Chitinase-3-like 1 regulating ERK-dependent PTX3 pathway

doi: 10.1186/s12974-020-02022-w

Figure Lengend Snippet: CHI3L1 mediates neuroinflammation through NF-κB pathway and ERK dependent PTX3 pathway

Article Snippet: Negative control (NC), PTX3 siRNA were purchased from OriGene Technologies, Inc. (Rockville, MD, USA). pcDNA3.1(+)-6 × Myc-CHI3L1 vector was cloned from Bionics (Seoul, Republic of Korea).

Techniques:

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: dePTX3 suppresses the growth and migration of human lung cancer cells. (A) CCK-8 assay was used to analyze the viability of A549 and SPCA1 cells. Cells were treated with or without rhPTX3 (100 ng/ml) or dePTX3 (rhPTX3; 100 ng/ml + PNGase F; 500 U/ml) for 0-72 h. Data were presented as a percentage of viable cells. (B) Lung cancer cells treated with TM (0.01, 0.05, 0.5 and 1 µ g/ml) for 48 h. (C) Immunofluorescent staining of PTX3 (red color) and N-glycan levels (green color) in A549 and SPCA1 cells analyzed following treatment with TM (0.5 µ g/ml) for 48 h. (D) SDS-PAGE gel (12%) stained with CBB and lectinblot against PHA to analyze N-glycans in lung cancer cells following treatment with TM for 48 h and PNGase F (500 U/ml) for 2 h of incubation. (E) Western blot analysis of dePTX3 in A549 and SPCA1 cells following treatment with TM or rhPTX3 + PNGase F. The lanes are labeled as follows: lane 1, glycosylated PTX3; lane 2, deglycosylated PTX3 by TM; lane 3, deglycosylated PTX3 by PNGase F. (F) Representative images of wound healing of A549 and SPCA1 cells analyzed after treatment with rhPTX3, dePTX3 and TM (0.5 µ g/ml), followed by measurement of relative wound width (48 h average wound width divided by 0 h wound width). (G) Microscopic images and graphs of transwell migration assay of A549 and SPCA1 cells following treatment with rhPTX3, dePTX3 and TM. Bar graph values represent the means ± SEM of 3 independent experiments. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Migration, CCK-8 Assay, Staining, Glycoproteomics, SDS Page, Incubation, Western Blot, Labeling, Transwell Migration Assay

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: PTX3 expression in human lung cancer samples and cell lines. (A) Immunohistochemical staining of PTX3 in paired human lung cancer tissue (P1, P2, P3) and normal lung tissue (N1, N2, N3) of the same patient (magnification, ×10). (B) PTX3 level in serum from lung cancer patients and normal healthy individuals by ELISA; * P<0.05. (C) PTX3 level in the supernatant collected from A549, SPCA1 and H1299 cells was examined by ELISA. (D) Immunofluorescent staining of PTX3 protein (red color) in A549, SPCA1 and H1299 lung cancer cells. DAPI (blue color) was used to stain the nuclei. (E) mRNA expression level of PTX3 detected by qPCR in A549, SPCA1 and H1299 cells. (F) PTX3 expression level in A549, SPCA1 and H1299 cells detected by western blot analysis. GAPDH was used as an internal control.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: PTX3 deglycosylation enhances the sensitivity of lung cancer cells to Cisplatin treatment. (A) suPTX3 level was detected by ELISA in the supernatant of A549 and SPCA1 cells following treatment with Cis (20 µ M) for 48 h. (B) Immunofluorescent staining of PTX3 (red color) in A549 and SPCA1 cells following treatment with Cis for 48 h. DAPI (blue color) was used to stain the nuclei. (C) Bar graph of A549 and SPCA1 cell viability analyzed following treatment with Cis and deglycosylated PTX3 (dePTX3). Each bar represents the average of 3 independent experiments. Data are expressed as a percentage of viable cells. (D) Western blot analysis of PCNA and AKT phosphorylation following treatment with Cis or dePTX3, and combined treatment for 48 h. (E) Western blot analysis for PTX3 expression in the cell lysate from control, mock and mutated PTX3 (mPTX3) in A549 and SPCA1 cells. (F) Western blot analysis of Cis, mPTX3 or Cis combined with mPTX3 and combined treatment for 48 h. The level of PCNA expression and AKT phosphorylation were examined by western blot analysis. GAPDH was used as an internal control. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Phospho-proteomics, Expressing, Control

Journal: International Journal of Oncology

Article Title: Tunicamycin enhances the suppressive effects of cisplatin on lung cancer growth through PTX3 glycosylation via AKT/NF-κB signaling pathway

doi: 10.3892/ijo.2018.4650

Figure Lengend Snippet: TM increases the sensitivity of lung cancer cells to cisplatin-induced apoptosis through deactivating AKT/NF-κB signaling pathway. (A) CCK-8 assay used to analyze the viability of A549 and SPCA1 cells treated with GDC0941 (4 µ M), IKK-16 (10 µ M), MEK162 (20 µ M) or rhPTX3 (100 ng/ml) for 48 h. (B) Western blot analysis of Bcl2 expression in A549 and SPCA1 cells, incubated with rhPTX3 (100 ng/ml) with or without NF-κB inhibitor IKK-16 (10 µ M). (C) A549 and SPCA1 cells were treated with TM (0.5 µ g/ml) + Cis (20 µ M) together or in combination. Western blot analysis of AKT, IKK, p65 phosphorylation in cytoplasmic and nuclear extracts of A549 and SPCA1 cells treated with TM, Cis or in combination. PARP was used as an internal control. (D) A549 and SPCA1 cells were double-stained with Annexin V-FITC/PI and analyzed by flow cytometry after 24 h of treatment with TM (0.5 µ g/ml), Cis (20 µ M) or combined treatment. The histogram showed the average percentage of total apoptotic cells in both A549 and SPCA1 cells. (E) Bcl2, Bax and cleaved PARP expression detected by western blot analysis following treatment with TM, Cis or in combination for 48 h. GAPDH was used as an internal control. Data represented the mean values ± SEM. * P<0.05 and ** P<0.01.

Article Snippet: Cell Counting kit-8 (CCK-8) was obtained from Beyotime Biotechnology (Shanghai, China); recombinant human PTX3 (rhPTX3) (#PROTP26022; Boster Biological Technology Co., Ltd., Fremont, CA); NF-κB inhibitor (IKK-16), PI3K inhibitor (GDC0941), MEK1/2 inhibitor, (MEK162) were purchased from Selleck Chemicals (Shanghai, China).

Techniques: CCK-8 Assay, Western Blot, Expressing, Incubation, Phospho-proteomics, Control, Staining, Flow Cytometry