Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Serial sections of normal and diseased pancreatic tissue were stained for CXCL12, CXCR4, CXCR7, and CK19. CXCL12 staining apparent in normal exocrine ducts was diminished in PDAC tissue. CXCR4 staining increased in PanIN and PDAC relative to normal ductal epithelium. CXCR7 expression was variable in normal epithelium, PanIN lesions, and PDAC. (A)Normal tissue from a single patient with healthy pancreas represents observations from 25 different normal tissues from 21 individual patients. (B)PDAC tissue from one patient represents observations from 82 different tissues from 29 different patients. 1000× magnification represents inset box at 200×. (C) Staining was quantified by blinded scoring of serial sections in relation to CK19 staining. (***) denotes P ≤0.001.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Staining, Expressing

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Representative 200× images of the same (A) normal or (B) pancreatic ductal adenocarcinoma (PDAC) tissues shown at 1000× magnification in . Representative serial section images of a pancreatic intestinal neoplasm (PanIN) lesion at (C) 200× or (D) 1000× magnification. Serial tissue sections were immunostained with antibodies to CK19, CXCL12, CXCR4, and CXCR7, or the isotype controls. n = 25 different normal tissues from 21 individual patients, 82 different tissues from 29 different PDAC patients, or 19 pathologically confirmed PanIN lesions.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques:

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: RT-PCR analysis revealed that (A) patient-derived pancreatic cancer cell lines (#1, #2, #3, #4) or (B) established cell lines lacked expression of CXCL12 and maintained expression of CXCR4. CXCR7 mRNA was present in 3 of 8 PDAC lines.Flow cytometric detection(C)of surface CXCR4 or CXCR7 protein expression. Cell lines treatedseven days with a concentration curve of (D) 5-aza-2-deoxycytidine (5-aza) restored expression of CXCL12.Linestreated 4 days with a concentration curve of (E) Trichostatin-A (TSA)restored CXCL12 mRNA expression in Capan2 cells.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Expressing, Concentration Assay

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: (A) Panc1 and MiaPaCa2 cells migrated towards exogenous gradients of CXCL12 in a range from 1 nM to 1000 nM under serum-free conditions(*), (**), and (***) denote P ≤0.05, P ≤0.01, and P ≤0.001, respectively, in comparison to unstimulated cells (NS). Receptor-null HPAFII cells did not migrate in response to CXCL12. (B) CXCL12 directs Panc1 cell chemoinvasion into a three-dimensional Matrigel plug. The positive control (+) was 10% serum-containing medium. (*), (**), and (***) denote P ≤0.05, P ≤0.01, and P ≤0.001, respectively, in comparison to unstimulated cells (NS).

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Positive Control

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Luciferase levels in control GFP or three different (31, #2, #3) CXCL12-expressing MiaPaCa2 clones as measured by spectrophotometer (A) or IVIS-100 biophotonic imager (B). Levels of CXCL12 were measured by ELISA (C) in established PDAC cell lines as well as GFP- and CXCL12-expressing MiaPaCa2-luciferase clones. (D) CXCL12-secreted by transfected MiaPaCa2-luciferase clones #1 and #2 stimulated U937 chemotaxis. Cells treated with neutralizing antibody to CXCL12 (αL12) confirmed the specificity of U937 chemotaxis. Values in A, C, and D are mean±SEM, n = 2–3.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Luciferase, Expressing, Clone Assay, Spectrophotometry, Enzyme-linked Immunosorbent Assay, Transfection, Chemotaxis Assay

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: (A) Transwell migration assays revealed significantly reduced chemotaxis of CXCL12-expressing clones (#1 and #2) compared to ligand null (GFP) cells. Attractants were serum-free media (--), 10% serum (+), or 10 nM CXCL12 (L12) in serum-free media. denote P ≤0.01 and P ≤0.001, respectively, compared to 10 nM CXCL12-stimulated GFP cells, n = 5. (B) CXCL12 re-expression diminished TGF-β (5 ng/mL)-induced chemotaxis relative to the CXCL12-null cells. (##) denotes P ≤0.01 compared to TGF-β-stimulated GFP cells, n = 4. (C) & (D) Representative images of experiments in (A) and (B) respectively. (E) CXCL12-expressing cells were significantly more adherent to tissue culture plastic compared to CXCL12-null cells. Untreated cells = (--), neutralizing antibody for CXCL12 activity = (αL12), and a positive control = 1 ng/mL of EGF (+). (##) denotes P ≤0.01 compared to 10% serum-stimulated control cells. (*), (**), and (***) denote P ≤0.05, P ≤0.01 and P ≤0.001, respectively, compared to unstimulated GFP cells, n = 7.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Migration, Chemotaxis Assay, Expressing, Clone Assay, Activity Assay, Positive Control

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: (A) Representative bioluminescence images of mice xenografted with GFP- or CXCL12-expressing cells at implantation (Day 0) or study endpoint (Day 28). (B) Whole-body in vivo radiance over time of GFP- and CXCL12-mice. (C) Ex vivo radiance of excised spleen (C), reflecting tumor cells at the site of injection, or the metastatic destination (D), reflecting decreased hepatic metastasis of CXCL12-expressing cells relative to GFP-cells. (**) denotes P ≤0.01, n = 4–5. (E) Representative H&E images showing pronounced tumor mass in the liver of control (GFP), relative to experimental (CXCL12) xenografted mice.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Expressing, In Vivo, Ex Vivo, Injection

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Apoptosis of GFP and CXCL12-expressing MiaPaCa2 cells were assessed using the caspase 3/7 glo assay.Cells were starved for 24 hours and cultured in 0% serum (A, C) or 1% serum (B, D) containing medium. (A–B) Apoptosis in adherent CXCL12-expressing cells was elevated compared to either GFP-expressing clones in serum-free conditions with no change seen in 1% serum. GFP-cells werestimulated with 100 µM gemcitabine as a control. (C–D) To measure cell number and detachment based apoptosis of cells in suspension, MiaPaCa2-luciferase cells were cultured on poly-HEMA. Using the Viacount reagent and flow cytometric cell counting, there was no difference in live cell number observed in non-adherent CXCL12-null (GFP) or expressing cells when cultured either in 0% (C) or 1% serum (D). Apoptosis of poly-HEMA cultured cells revealed an in increase in active caspase-3/7 restricted to cells cultured in serum-free conditions only (C) with no change observed in 1% serum (D). Gemcitabine (GEM) was used as a control for decreased cell count and increased apoptosis.(*), (**), and (***) denote P ≤0.05, P ≤0.01, and P ≤0.001 respectively in comparison to control cells (GFP). Values are mean±SEM, n = 4–5.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Expressing, Glo Assay, Cell Culture, Clone Assay, Luciferase, Cell Counting

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Population growth of adherent GFP and CXCL12-expressing MiaPaCa2 cells was assessed using the Viacount reagent and flow cytometric cell counting (A–B). CXCL12-expressing cells starved for 24 hours and cultured in both 0% serum (A) or 1% serum (B) containing medium were found to have decreased population growth.(B) Two CXCL12-expressing clones (#1, #2) were compared to GFP alone or GFP + gemcitabine (GEM) controls in 1% serum containing medium. Doubling time of adherent clones (T 2 ) was calculated using a linear regression of the data to determine slope and the intercept at y = 10 6 , with increased T 2 observed in both CXCL12-expressing clones. (C–D) Propidium Iodide cell cycle analysis revealed a decrease in percentage of cells in the G 2 phase in CXCL12-expressing cells compared to GFP controls. (*), (**), and (***) denote P ≤0.05, P ≤0.01, and P ≤0.001 respectively in comparison to control cells (GFP). Values are mean±SEM, n = 4–5.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Expressing, Cell Counting, Cell Culture, Clone Assay, Cell Cycle Assay

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: (A) Kaplan-Meier survival curves for GFP- and CXCL12-expressing cell groups. Three experimental CXCL12 PDAC injected mice were removed for non-study reasons (tick marks). (B) Percent change in bioluminescence from baseline-level measured at day 7 for both GFP and CXCL12 engrafted mice. Dotted lines represent individual mice. Solid lines are quadratic regression fitted curves of each group. Statistical comparison was done between both groups independent of time. (C–D) Representative bioluminescence images of mice from each groupat days 7, 49, and endpoint for GFP (98) or CXCL12 (106). (E) Tumor wet weight was significantly reduced in CXCL12-expressing tumors relative GFP-tumors. Representative photomicrographs are shown in lower panels. (F) CXCL12-producing tumors had significantly fewer Ki-67 positive cells compared to GFP-expressing tumors, as counted in a cross-section of each tumor normalized to the total cross-sectional area of each tumor. (G) Representative images of Ki-67immunostaining and rabbit isotype control (inset, Rab IgG) are shown.n = 8–10. (**) and (***) denote P ≤0.01 and P ≤0.001 respectively, between CXCL12-expressing and control xenografted mice.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Expressing, Injection

Journal: PLoS ONE

Article Title: CXCL12 Chemokine Expression Suppresses Human Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0090400

Figure Lengend Snippet: Ex vivo bioluminescence analysis revealed significantly decreased metastasis to the liver (A), lung (C), and mesenteric lymph nodes (D) of CXCL12-expressing cells compared to GFP-controls. Representative biophotonic images of hepatic metastases are shown in panel B. (**) and (***) denote statistically significant P ≤0.01 and P ≤0.001, respectively, differences between CXCL12-expressing and control tumor engrafted mice. n = 8–10 mice in each group.

Article Snippet: Secreted CXCL12 protein from supernatant of pancreatic cancer cells cultured in serum-free media was detected by ourpreviously established sandwich ELISA method using antibodies from R&D Systems (monoclonal mouse and human CXCL12 (MAB350) and goat anti-human CXCL12 (BAF310) .Cell surface CXCR4 or CXCR7was detected using the aforementioned antibodies (Abcam) along withFITC-conjugated secondary antibodies using our previously established method .

Techniques: Ex Vivo, Expressing

Journal: Journal of Applied Oral Science

Article Title: CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS

doi: 10.1590/1678-7757201300004

Figure Lengend Snippet: Production of chemokines by different subtypes of dental pulp fibroblasts. Cultured dental pulp fibroblasts from permanent (n=3) and deciduous (n=2) teeth were stimulated by P. gingivalis LPS at the indicated concentrations (n=6). Cell supernatants were collected after 1 (A and D), 6 (B and E) and 24 (C and F) h. Production of CCL3 (A, B and C) and CXCL12 (D, E and F) was detected by means of ELISA. (*) p<0.05; (**) p<0.01 and (***) p<0.001 in comparison with culture medium alone (0). (##) p<0.01 and (###) p<0.001 in comparison with the other cellular subtype at the same experimental condition.

Article Snippet: The production of CCL3 and CXCL12 was detected by means of enzyme-linked immunosorbent assay (ELISA) (Anti-human CCL3/MIP-1a Antibody - AF-270-NA and Biotinylated Anti-human CCL3/MIP-1a Antibody - BAF 270, Monoclonal Anti-human/mouse CXCL12/SDF-1 Antibody - MAB 350 and Biotinylated Anti-human CXCL12/SDF-1 Antibody - BAF 310, R&D Systems, Minneapolis, USA, respectively) according to the manufacturer's instructions.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Comparison between total Allred scores and mRNA levels of CXCR4, CXCR7 and CXCL12 in primary sites and metastatic lymph nodes. A,C,E, Comparison of total Allred scores of CXCR4, CXCR7 and CXCL12 in primary site (PS) between low and high groups. B,D,F, Comparison of total Allred scores of CXCR4, CXCR7 and CXCL12 in metastatic lymph nodes (MLN) between low and high groups. C t , threshold cycle. Δ C t , C t of target − C t of reference (β‐actin). The statistical difference was assessed by two‐way analysis of variance. * P < .05, ** P < .01, *** P < .0001

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Comparison

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Expression of CXCR4, CXCR7 and CXCL12 in primary sites (PS) and metastatic lymph nodes (MLN). A, CXCR4 immunostaining in lung tumor cells. IS = 0 (left) and IS = 3 (right) (×200 and ×100, respectively). B, CXCR7 immunostaining in lung tumor cells. IS = 0 (left) and IS = 3 (right) (×200 and ×400, respectively). C, CXCL12 immunostaining in lung tumor cells. IS = 0 (left) and IS = 3 (right) (×200 and ×100, respectively). D, The bar graphs show the percentage of expression of each protein in PS or MLN. White bar: low expression group; black bar: high expression group. IS, intensity score; MLN, metastatic lymph node; PS, primary site

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Expressing, Immunostaining

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Comparison of CXCL12 total Allred score grouped by cut‐off in stromal cells and tumor cells in metastatic lymph nodes. The bar graphs show the distribution of high and low CXCL12 protein expression grouped by cut‐off in stromal and tumor cells in metastatic lymph nodes (MLN). White bar: low CXCL12 protein expression group; black bar: high CXCL12 protein expression group. *** P < .0001

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Comparison, Expressing

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Postoperative overall survival curves of patients with non‐small cell lung cancer (NSCLC) in the primary site and metastatic lymph nodes. A, Kaplan‐Meier curves show overall survival according to CXCR4 in the primary site (PS). Patients with high CXCR4 expression had a poorer prognosis than patients with low CXCR4 expression ( P < .05, log‐rank test). B, Kaplan‐Meier curves show overall survival according to CXCL12 expressions in metastatic lymph nodes (MLN). Patients with high CXCL12 expression had a poorer prognosis than patients with low CXCL12 expression ( P < .05, log‐rank test). C, Subgroup analysis using both CXCR4 expression in PS and CXCL12 expression in MLN. Patients with both high CXCR4 and high CXCL12 expression had a poorer prognosis than the other patient groups ( P < .005)

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Expressing

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Tumor cells and cancer‐associated fibroblasts produce increased CXCL12 protein in primary sites and metastatic lymph nodes. A, Immunostaining of primary site of invasive human lung cancer for CXCL12 and α‐SMA (×400). B, Immunostaining of metastatic lymph node of invasive human lung cancer for CXCL12 and α‐SMA (×400). DAPI (blue) indicates cell nuclei (upper left), α‐SMA (green, upper right), CXCL12 (red, lower left) and merge (lower right). *Tumor cluster

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Immunostaining

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Clinicopathological characteristics of patients according to CXCR7 and CXCL12 protein expression in metastatic lymph nodes

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Expressing

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Correlations between CXCR4 expression in the primary site and CXCR7 and CXCL12 expression in the metastatic lymph nodes

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Expressing

Journal: Cancer Science

Article Title: Correlation between CXCR4/CXCR7/CXCL12 chemokine axis expression and prognosis in lymph‐node‐positive lung cancer patients

doi: 10.1111/cas.13422

Figure Lengend Snippet: Univariate and multivariate analysis of the relationships between overall survival and the clinical factors of the NSCLC patients

Article Snippet: Samples were incubated with Blocking One Histo (Nakalai Tesque, Kyoto, Japan) to eliminate nonspecific binding, and then incubated overnight at 4°C with primary antibodies: α‐SMA (monoclonal rabbit anti‐human ACTA2/Smooth Muscle Actin; LifeSpan BioSciences, Seattle, WA, USA, 1:200) or CXCL12 (polyclonal goat anti‐human SDF‐1; Gene‐Tex, Irvine, CA, USA; 1:200).

Techniques: Expressing

Journal: International journal of cancer

Article Title: Locally generated VGVAPG and VAPG elastin-derived peptides amplify melanoma invasion via the galectin-3 receptor.

doi: 10.1002/ijc.23296

Figure Lengend Snippet: FIGURE 4 – Elastin-derived peptides increase attachment and the expression of major adhesion molecules. (A) The attachment of melanoma cells toward VGVAPG, VAPG and scrambled peptides was quantified by ELISA reader after crystal violet staining and plotted by the OD at a wavelength of 560 nm. (B) The expression of CD44 following the peptide treatments (100 lg/ml) in WM35 (a) and in HT168-M1 (b) cells was determined by flow cytometry; on the histogram plot the black bold line represents the secondary antibody (left); dotted line represents control cells; gray bold line represents VAPG treatment; broken line represents VGVAPG treatment (right); the X Geo Mean value was determined by flow cytometry (c) and the relative quantity of CD44 mRNA level was determined by real-time PCR (d). (C) The expression of ICAM-1 follow- ing the peptide treatments (100 lg/ml) in WM35 (a) and in HT168-M1 (b) cells was determined by flow cytometry; on the histogram plot the black bold line represents the secondary antibody (left); dotted line represents control cells; gray bold line represents VAPG treatment; broken line represents VGVAPG treatment (right); the X Geo Mean value was determined by flow cytometry (c) and the relative quantity of ICAM-1 mRNA level was determined by real-time PCR (d). (D) The expression of NCAM following the peptide treatments (100 lg/ml) in WM35 (a) and in HT168-M1 (b) cells was determined by flow cytometry; on the histogram plot the black bold line represents the secondary antibody (left); dotted line represents control cells; gray bold line represents VAPG treatment; broken line represents VGVAPG treatment (right); the X Geo Mean value was determined by flow cytometry (c) and the relative quantity of NCAM mRNA level was determined by real-time PCR (d). Results represent the mean 6 SEM of triplicate experiments; *p < 0.05; **p < 0.01 when compared to levels in the relevant controls.

Article Snippet: The following primary antibodies were used: mouse antihuman MMP-2 and MMP-3 (1:1,000; Chemicon), mouse antihuman CD44, ICAM-1 and NCAM (1:1,000; R&D Systems), goat antihuman VEGF-C and CXCL-12 (1:2,000; Sigma–Aldrich), mouse antihuman CXCR-4 (1:1,000; Sigma–Aldrich) and rat antitubulin (0.5 lg/ml, Serotec, Oxford, UK).

Techniques: Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Cytometry, Control, Real-time Polymerase Chain Reaction