Journal: Cancer research

Article Title: Proinflammatory CXCL12-CXCR4/CXCR7 signaling axis drives Myc-induced prostate cancer in obese mice

doi: 10.1158/0008-5472.CAN-17-0284

Figure Lengend Snippet: A, Increased mRNA expression of CXCL12, CXCR4 and CXCR7 in the SVF of obese HiMyc mice. mRNA was isolated from the SVF of adipose tissue surrounding the ventral prostate of HiMyc mice at 6 months of age. Fold change in gene expression in obese mice fed the DIO (60kcal% fat) diet relative to mice fed the control (10kcal% fat) diet is shown. Quantitation of fold change of mRNA is presented as mean ± SEM of three independent experiments. Significantly different, *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 (Student’s t-test). B,C, Immunofluorescence analysis of representative prostate tissue sections from age-matched HiMyc mice fed on CR, Control or DIO diet with antibodies against CXCL12, α-SMA, CXCR4 and CXCR7 with red and green fluorophore-conjugated secondary antibodies. Shown are representative images from one mouse out of a group of 5 mice. All tissue sections from each diet group gave similar results. Scale bar 100 μm.

Article Snippet: The samples were then blocked for 1h at room temperature and incubated with primary antibodies [Perilipin (9349) from Cell Signaling, CD3 (550275) from BD Pharmingen, RM0029-11H3 (ab56297), CXCR7 (ab72100) from Abcam, CXCL12 (sc-28876) from Santa Cruz, CXCR4 (MAB21651) from R&D Systems and αSMA (C6198) from Sigma] overnight at 4°C.

Techniques: Expressing, Isolation, Quantitation Assay, Immunofluorescence

Journal: Cancer research

Article Title: Proinflammatory CXCL12-CXCR4/CXCR7 signaling axis drives Myc-induced prostate cancer in obese mice

doi: 10.1158/0008-5472.CAN-17-0284

Figure Lengend Snippet: CXCL12 induces migration and invasion of HMVP2 cells. A, B. cells in 12 well plates scratched with 200 μl pipette tips and treated with vehicle control, CXCL12 (100 ng/mL), IL-6 (10 ng/mL), AMD3100 (40 ug/mL) or CXCL12 + AMD3100 at 0 and 18 h post-treatment. Quantitation of migration levels are presented as mean ± SEM of three independent experiments. ***P<0.001; (One Way ANOVA). C, D. NMVP and HMVP2 cells were seeded in Matrigel in the upper transwell chambers and then placed into 24 well plates with medium that contained the vehicle (control), CXCL12 (100 ng/mL), IL-6 (10 ng/mL), AMD3100 (40 ug/ml) or CXCL12 + AMD3100. After 18h, the invading cells in the lower chamber were fixed, stained and visualized by light microscopy. Quantitation of invading cells presented as mean ± SEM of three independent experiments. ***P<0.001; (One Way ANOVA)

Article Snippet: The samples were then blocked for 1h at room temperature and incubated with primary antibodies [Perilipin (9349) from Cell Signaling, CD3 (550275) from BD Pharmingen, RM0029-11H3 (ab56297), CXCR7 (ab72100) from Abcam, CXCL12 (sc-28876) from Santa Cruz, CXCR4 (MAB21651) from R&D Systems and αSMA (C6198) from Sigma] overnight at 4°C.

Techniques: Migration, Transferring, Quantitation Assay, Staining, Light Microscopy

Journal: Cancer research

Article Title: Proinflammatory CXCL12-CXCR4/CXCR7 signaling axis drives Myc-induced prostate cancer in obese mice

doi: 10.1158/0008-5472.CAN-17-0284

Figure Lengend Snippet: CXCR4 and CXCR7 knockdown decreases migration and invasion in HMVP2 cells. Cells (A, transfected with control or CXCR4 shRNA; C, transfected with control or CXCR7 shRNA) in 12 well plates scratched with 200 μl pipette tips and treated with vehicle control or CXCL12 (100 ng/mL). Quantitation of migration levels were presented as mean ± SEM of three independent experiments. ***P<0.001; (Student’s t-test). Cells (B, transfected with control or CXCR4 shRNA; D, transfected with control or CXCR7 shRNA) were seeded in Matrigel in the upper transwell chambers and then placed into 24 well plates with medium that contained the vehicle (control) or CXCL12 (100 ng/mL). After 18h, the invading cells in the lower chamber were fixed, stained and visualized by light microscopy. Quantitation of invading cells were presented as mean ± SEM of three independent experiments. ***P<0.001; (Student’s t-test)

Article Snippet: The samples were then blocked for 1h at room temperature and incubated with primary antibodies [Perilipin (9349) from Cell Signaling, CD3 (550275) from BD Pharmingen, RM0029-11H3 (ab56297), CXCR7 (ab72100) from Abcam, CXCL12 (sc-28876) from Santa Cruz, CXCR4 (MAB21651) from R&D Systems and αSMA (C6198) from Sigma] overnight at 4°C.

Techniques: Migration, Transfection, shRNA, Transferring, Quantitation Assay, Staining, Light Microscopy

Journal: Cancer research

Article Title: Proinflammatory CXCL12-CXCR4/CXCR7 signaling axis drives Myc-induced prostate cancer in obese mice

doi: 10.1158/0008-5472.CAN-17-0284

Figure Lengend Snippet: Activation of NFκB, STAT3 and MAPK signaling by CXCL12 in HMVP2 cells. A,B. Protein lysates were prepared from cultured NMVP and HMVP2 cells treated with CXCL12 (100 ng/mL) for 5 to 30 min and subjected to Western blot analysis using antibodies for the indicated phospho and total proteins. Relative protein levels were quantitated by densitometry; β-actin was used to control for protein loading. Representative blots of three separate experiments with similar results are shown.

Article Snippet: The samples were then blocked for 1h at room temperature and incubated with primary antibodies [Perilipin (9349) from Cell Signaling, CD3 (550275) from BD Pharmingen, RM0029-11H3 (ab56297), CXCR7 (ab72100) from Abcam, CXCL12 (sc-28876) from Santa Cruz, CXCR4 (MAB21651) from R&D Systems and αSMA (C6198) from Sigma] overnight at 4°C.

Techniques: Activation Assay, Cell Culture, Western Blot

Journal: Chinese Journal of Cancer

Article Title: The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

doi: 10.1186/s40880-017-0246-z

Figure Lengend Snippet: Relationships between CXCR4, CXCL12, and Cyclin D1 expression levels and clinicopathologic characteristics of sporadic MPNST patients

Article Snippet: Immunohistochemical (IHC) staining of TMAs was performed using the streptavidin–peroxidase method [ ] with the following biotinylated primary antibodies: anti-CXCR4 antibody (1:100, ab124824, Abcam, Cambridge, UK), anti-CXCL12 antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Cyclin D1 antibody (1:50, Beijing Zhong Shan Golden Bridge Biotechnology Co. Ltd, Beijing, China).

Techniques: Expressing

Journal: Chinese Journal of Cancer

Article Title: The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

doi: 10.1186/s40880-017-0246-z

Figure Lengend Snippet: Immunohistochemical analysis of C-X-C motif chemokine receptor 4 (CXCR4), C-X-C motif chemokine ligand 12 (CXCL12), and Cyclin D1 protein expression in sporadic malignant peripheral nerve sheath tumor tissue samples. a CXCR4 is mainly expressed in the cytoplasm and membrane. b CXCL12 is mainly presented in the cytoplasm and plasma membrane. c Cyclin D1 is expressed in the nucleus

Article Snippet: Immunohistochemical (IHC) staining of TMAs was performed using the streptavidin–peroxidase method [ ] with the following biotinylated primary antibodies: anti-CXCR4 antibody (1:100, ab124824, Abcam, Cambridge, UK), anti-CXCL12 antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Cyclin D1 antibody (1:50, Beijing Zhong Shan Golden Bridge Biotechnology Co. Ltd, Beijing, China).

Techniques: Immunohistochemical staining, Expressing, Membrane, Clinical Proteomics

Journal: Chinese Journal of Cancer

Article Title: The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

doi: 10.1186/s40880-017-0246-z

Figure Lengend Snippet: Prognostic values of CXCR4, CXCL12, Cyclin D1 expression, tumor recurrence, and tumor metastasis in sporadic malignant peripheral nerve sheath tumor patients. a No association is shown between CXCR4 expression and disease-free survival (DFS) ( P > 0.05). b Overall survival (OS) is longer for patients with high CXCR4 expression than for those with low CXCR4 expression ( P < 0.05). c No association is shown between CXCL12 expression and DFS ( P > 0.05). d No association is shown between CXCL12 expression and OS ( P > 0.05). e No association is shown between Cyclin D1 expression and DFS ( P > 0.05). f No association is shown between Cyclin D1 expression and OS ( P > 0.05). g OS is shorter for patients with recurrence than for those without recurrence ( P < 0.01). h OS is shorter for patients with disease metastasis than for those without metastasis ( P < 0.01)

Article Snippet: Immunohistochemical (IHC) staining of TMAs was performed using the streptavidin–peroxidase method [ ] with the following biotinylated primary antibodies: anti-CXCR4 antibody (1:100, ab124824, Abcam, Cambridge, UK), anti-CXCL12 antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Cyclin D1 antibody (1:50, Beijing Zhong Shan Golden Bridge Biotechnology Co. Ltd, Beijing, China).

Techniques: Expressing

Journal: Chinese Journal of Cancer

Article Title: The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

doi: 10.1186/s40880-017-0246-z

Figure Lengend Snippet: Univariate analysis of the prognostic values of clinicopathologic characteristics and protein expression in sporadic MPNST patients

Article Snippet: Immunohistochemical (IHC) staining of TMAs was performed using the streptavidin–peroxidase method [ ] with the following biotinylated primary antibodies: anti-CXCR4 antibody (1:100, ab124824, Abcam, Cambridge, UK), anti-CXCL12 antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-Cyclin D1 antibody (1:50, Beijing Zhong Shan Golden Bridge Biotechnology Co. Ltd, Beijing, China).

Techniques: Expressing

Journal: Journal of Neuroinflammation

Article Title: CXCL12 in astrocytes contributes to bone cancer pain through CXCR4-mediated neuronal sensitization and glial activation in rat spinal cord

doi: 10.1186/1742-2094-11-75

Figure Lengend Snippet: Expression and distribution of CXCL12 protein in the spinal cord after TCI in rats. (A) Western blot analysis shows time course of CXCL12 expression in sham and TCI rats. Five spinal cord segments were included in each group. Representative bands are shown on the top; data summary is shown on the bottom. * P < 0.05, ** P < 0.01 versus sham control. (B) Immunofluorescence shows expression of CXCL12 protein (green) in rat spinal cord at different time points. Tissues were collected at day 3, 5, 10, 14 after TCI. Original magnification: 200× for all the confocal images showing the dorsal horn ipsilateral to TCI (bar = 100 μm).

Article Snippet: The sections were first blocked with 5% donkey serum and 0.3% Triton X-100 for 1hour at room temperature, then incubated overnight at 4°C with the following primary antibodies: rabbit anti-CXCL12 polyclonal antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, California, United States), rabbit anti-CXCR4 polyclonal antibody (1:200, ab2074, Abcam, Cambridge, Massachusetts, United States), mouse anti-NeuN monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, MAB377X, Millipore, Billerica, Massachusetts, United States), mouse anti-GFAP monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, #3655, Cell Signaling Technology, Beverly, Massachusetts, United States), goat anti-IBA1 polyclonal antibody (1:400, ab5076, Abcam, Cambridge, Massachusetts, United States ), rabbit anti-c-Fos polyclonal antibody (1:800, ab7963, Abcam, Cambridge, Massachusetts, United States ).

Techniques: Expressing, Western Blot, Control, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: CXCL12 in astrocytes contributes to bone cancer pain through CXCR4-mediated neuronal sensitization and glial activation in rat spinal cord

doi: 10.1186/1742-2094-11-75

Figure Lengend Snippet: Cellular localization of CXCL12 expression in spinal cord dorsal horn of sham and TCI rats. Double staining shows that CXCL12 (red) is colocalized with GFAP (green, middle), a marker for astrocytes, but not with NeuN (green, left), a marker for neurons or IBA1 (green, right), a marker for microglia. Tissues were collected 10 days after TCI. Two single-stained images were merged. Original magnification: 400× for all the confocal images showing the dorsal horn ipsilateral to TCI (bar = 50 μm).

Article Snippet: The sections were first blocked with 5% donkey serum and 0.3% Triton X-100 for 1hour at room temperature, then incubated overnight at 4°C with the following primary antibodies: rabbit anti-CXCL12 polyclonal antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, California, United States), rabbit anti-CXCR4 polyclonal antibody (1:200, ab2074, Abcam, Cambridge, Massachusetts, United States), mouse anti-NeuN monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, MAB377X, Millipore, Billerica, Massachusetts, United States), mouse anti-GFAP monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, #3655, Cell Signaling Technology, Beverly, Massachusetts, United States), goat anti-IBA1 polyclonal antibody (1:400, ab5076, Abcam, Cambridge, Massachusetts, United States ), rabbit anti-c-Fos polyclonal antibody (1:800, ab7963, Abcam, Cambridge, Massachusetts, United States ).

Techniques: Expressing, Double Staining, Marker, Staining

Journal: Journal of Neuroinflammation

Article Title: CXCL12 in astrocytes contributes to bone cancer pain through CXCR4-mediated neuronal sensitization and glial activation in rat spinal cord

doi: 10.1186/1742-2094-11-75

Figure Lengend Snippet: Expression of CXCL12 protein in DRG after TCI in rats. (A) Western blot analysis shows time course of DRG CXCL12 expression in sham and TCI rats. (B) Western blot analysis shows DRG or SC CXCL12 expression in sham and TCI rats at day 1. Five samples were included in each group. Representative bands are shown on the top; data summary is shown on the bottom. ** P < 0.01 versus sham control.

Article Snippet: The sections were first blocked with 5% donkey serum and 0.3% Triton X-100 for 1hour at room temperature, then incubated overnight at 4°C with the following primary antibodies: rabbit anti-CXCL12 polyclonal antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, California, United States), rabbit anti-CXCR4 polyclonal antibody (1:200, ab2074, Abcam, Cambridge, Massachusetts, United States), mouse anti-NeuN monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, MAB377X, Millipore, Billerica, Massachusetts, United States), mouse anti-GFAP monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, #3655, Cell Signaling Technology, Beverly, Massachusetts, United States), goat anti-IBA1 polyclonal antibody (1:400, ab5076, Abcam, Cambridge, Massachusetts, United States ), rabbit anti-c-Fos polyclonal antibody (1:800, ab7963, Abcam, Cambridge, Massachusetts, United States ).

Techniques: Expressing, Western Blot, Control

Journal: Journal of Neuroinflammation

Article Title: CXCL12 in astrocytes contributes to bone cancer pain through CXCR4-mediated neuronal sensitization and glial activation in rat spinal cord

doi: 10.1186/1742-2094-11-75

Figure Lengend Snippet: Intrathecal administration of fluorocitrate or SP600125 reverses TCI-induced upregulation of CXCL12 protein in the spinal cord. (A) Confocal images show inhibitory effect of fluorocitrate on induction of astrocytic marker GFAP (green) after TCI. Original magnification: ×200. (B) Mean green immunofluorescence intensity of GFAP of A among sham, TCI and TCI + Fluorocitrate group. For control experiments, saline was used. Fluorocitrate (1 nmol/10 μl, i.t.) or saline (10 μl, i.t.) was given once a day on postoperative days 8, 9 and 10, respectively. Tissues were collected 4 hours after the last spinal injection (n = 5 in each group). * P < 0.05, ** P < 0.01 versus sham + saline. ## P < 0.01 versus TCI + saline. (C) Western blot and data summary show inhibitory effect of fluorocitrate on TCI-induced increased expression of CXCL12 protein (n = 4 in each group). (D) Western blot and data summary show inhibitory effect of SP600125 on TCI-induced increased expression of CXCL12 protein (n = 4 in each group). SP600125 (20 nmol/10 μl, i.t.) or DMSO (for control, 10 μl, i.t.) was given once a day on postoperative days 8, 9 and 10, respectively. Tissues were collected 6 hours after the last spinal injection. ** P < 0.01 versus sham + DMSO. ## P < 0.01 versus TCI + DMSO.

Article Snippet: The sections were first blocked with 5% donkey serum and 0.3% Triton X-100 for 1hour at room temperature, then incubated overnight at 4°C with the following primary antibodies: rabbit anti-CXCL12 polyclonal antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, California, United States), rabbit anti-CXCR4 polyclonal antibody (1:200, ab2074, Abcam, Cambridge, Massachusetts, United States), mouse anti-NeuN monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, MAB377X, Millipore, Billerica, Massachusetts, United States), mouse anti-GFAP monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, #3655, Cell Signaling Technology, Beverly, Massachusetts, United States), goat anti-IBA1 polyclonal antibody (1:400, ab5076, Abcam, Cambridge, Massachusetts, United States ), rabbit anti-c-Fos polyclonal antibody (1:800, ab7963, Abcam, Cambridge, Massachusetts, United States ).

Techniques: Marker, Immunofluorescence, Control, Saline, Injection, Western Blot, Expressing

Journal: Journal of Neuroinflammation

Article Title: CXCL12 in astrocytes contributes to bone cancer pain through CXCR4-mediated neuronal sensitization and glial activation in rat spinal cord

doi: 10.1186/1742-2094-11-75

Figure Lengend Snippet: Effects of CXCL12 neutralizing antibody in spinal cord on mechanical and thermal hypersensitivity induced by TCI. (A, B) Intrathecal injection of CXCL12 neutralizing antibody (10 μg, but not 1 μg) partially and transiently reverses TCI-induced mechanical allodynia (A) and thermal hyperalgesia (B) in the feet ipsilateral to TCI. Pain behaviors were measured on postoperative day 10. # P < 0.05, ## P < 0.01 versus TCI + control IgG. (C, D) Intraspinal injection of CXCL12 neutralizing antibody (10 μg/10 μl, once a day for three consecutive days on days 3, 4, and 5 after TCI) delays TCI-induced induction of mechanical allodynia (C) and heat hyperalgesia (D) . ** P < 0.01 versus sham + control IgG. # P < 0.05, ## P < 0.01 versus TCI + control IgG. TCI was performed on day 0. The dose of control IgG was 10 μg/10 μl. Each administration is indicated by an arrow on the corresponding time point. Ten rats were included in each group.

Article Snippet: The sections were first blocked with 5% donkey serum and 0.3% Triton X-100 for 1hour at room temperature, then incubated overnight at 4°C with the following primary antibodies: rabbit anti-CXCL12 polyclonal antibody (1:100, sc-28876, Santa Cruz Biotechnology, Santa Cruz, California, United States), rabbit anti-CXCR4 polyclonal antibody (1:200, ab2074, Abcam, Cambridge, Massachusetts, United States), mouse anti-NeuN monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, MAB377X, Millipore, Billerica, Massachusetts, United States), mouse anti-GFAP monoclonal antibody (1:400, Alexa Fluor 488 Conjugate, #3655, Cell Signaling Technology, Beverly, Massachusetts, United States), goat anti-IBA1 polyclonal antibody (1:400, ab5076, Abcam, Cambridge, Massachusetts, United States ), rabbit anti-c-Fos polyclonal antibody (1:800, ab7963, Abcam, Cambridge, Massachusetts, United States ).

Techniques: Injection, Control