Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 1. Expression levels of DDR2 transcripts and proteins in HNSCC clinical specimens. (A) Expression levels of DDR2 in 29 pairs of fresh primary HNSCC and corresponding normal clinical specimens were measured by qPCR. One of the low grade tumors’ transcripts (number 5) was used for normalization. (B) Immunohistochemical staining for DDR2 expression in 79 human HNSCC specimens (54 specimens of low grade tumors and 25 specimens of high grade tumors) and 39 cancer-associated normal (cancer-assoc. nl) specimens. Shown is a representative example of high grade HNSCC sample (right), low grade HNSCC sample (middle) and cancer-associated normal tissue sample (left). (C) Staining evaluation for the immunohistochemical staining of DDR2 expression. The staining grade is stratified as absent (−), weak (+), moderate (++), or strong (+++). Shown is a representative example of each grade. (D) Statistical analysis for immunohistochemical staining of DDR2 expression between human HNSCC specimens and cancer-associated normal specimens. (E) Statistical analysis for the relationship between the immunohistochemical staining of DDR2 expression and HNSCC characteristics. Data are presented as the mean ± SD or n (number of samples). Statistical significance was evaluated with the Student t test or χ2 test.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 2. Expression of DDR2, E-cadherin, and vimentin in HNSCC cell lines. (A and B) Tca8113, TB, SCC25, and FaDu cells were subjected to analysis of mRNA and protein expression of E-cadherin and vimentin by qPCR and western blot, respectively. Statistical significance was evaluated with the Student t test. Results display the mean ± SD. ***P < 0.001. (C) mRNA and protein expression of DDR2 in HNSCC cell lines was analyzed by reverse transcription PCR and western blot, respectively. β-actin was used as a loading control.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: Expressing, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 3. DDR2 overexpression has no effect on cell proliferation in HNSCC cells. (A) Tca8113 and FaDu cells (parental, EGFP-, and DDR2-transduced) were cultured in serum-free medium for 24 h. Thereafter, protein and RNA from these cells were extracted and analyzed for DDR2 expression using western blot and qPCR. (B) The cell growth curves of Tca8113 cells (parental, EGFP-, and DDR2-transduced) by MTT method. (C) Tca8113 cells (parental, EGFP-, and DDR2-transduced) treated by EdU were viewed and photographed using a fluorescence microscope. The staining positive rate was counted as positive cells/overall cells × 100%. For each group, ten random high-power fields (400×) were chosen and the cell number was counted at least three times. The histogram represents the staining positive rate of each group. (D) Tca8113 cells infected with Ad-DDR2 were similar to the controls by FAC assays. Statistical significance was evaluated with the Student t test. Results display the mean ± SD. ***P < 0.001.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: Over Expression, Cell Culture, Expressing, Western Blot, Fluorescence, Microscopy, Staining, Infection

Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 4. Overexpression of DDR2 enhances the invasion and migration in HNSCC cells. (A and B) Tca8113 and FaDu cells (parental, EGFP-, and DDR2-transduced) were incubated in serum-free medium with or without 2 µg/mL type I collagen (Col I) for 24 h. The histogram represents the average number of invaded and migrated cells in five random low-power fields (200×). The numbers from the control groups were used as control. Statistical significance was evaluated with the Student t test. Results display the mean ± SD. ***P < 0.001. (C) The cells in (A) were subjected to analysis of protein expression of MMP-2 and MMP-9 using western blot.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: Over Expression, Migration, Incubation, Expressing, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 5. Overexpression of DDR2 accelerates the process of hypoxia-induced EMT in HNSCC cells. (A) Tca8113 and FaDu cells (parental and DDR2-transduced) were cultured under normoxic or hypoxic condition for 48 h, and the morphology change of cells were photographed in low-power field (200×). (B) The cells in (A) were stained for the expression of E-cadherin (red) and vimentin (green) by immunocytofluorescence. Nuclei are labeled by DAPI (blue). Scale bar 10 µm. (C) The cells in (A) were used for immunoblot analysis of E-cadherin and vimentin expression.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: Over Expression, Cell Culture, Staining, Expressing, Labeling, Western Blot

Journal: Cancer Biology & Therapy

Article Title: Overexpression of DDR2 contributes to cell invasion and migration in head and neck squamous cell carcinoma

doi: 10.4161/cbt.28181

Figure Lengend Snippet: Figure 6. DDR2 enhances the metastasis potential of HNSCC cells in vivo. Tca8113 cells (parental, EGFP-, and DDR2-transduced) (2 × 106/200 μL PBS) were simultaneously injected via the tail veins and subcutaneously into the hind legs of the 6-wk-old female nude mice. At day 42, mice were killed and the lungs were fixed in 4% formaldehyde for 24 h. (A) Tumor growth curve. The tumor growth was assessed every 3 d until day 42 by measuring two perpendicular diameters and calculating the volume in mm3. The typical photographs of tumors are shown. (B) The white colonies on the lung surface were enumerated macroscopically. Black arrows indicate lung metastatic tumors. Colonies were enumerated and the relative increases compared with the control groups were calculated. (C) Representative lung tissue sections from each group are shown (H&E; original magnification 100×). Black arrows indicate lung metastatic tumors. The number of lung metastatic foci in each group (n = 4) was calculated. (D and E) Immunostaining of MMP-2 and MMP-9 were performed on lung metastatic tumors (original magnification 200× and 400×). Shown are representative examples from EGFP- and DDR2-transduced groups. Statistical significance was evaluated with the Student t test. Results display the mean ± SD .***P < 0.001.

Article Snippet: Anti-DDR2 (MMB2538 and BAF2538) antibodies were purchased from R&D Corporation (R&D), anti-E-cadherin (ab11512, Abcam), anti-vimentin, anti-β-actin, anti-MMP-2, and anti-MMP-9 (Cell Signaling).

Techniques: In Vivo, Injection, Immunostaining

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 1. Interleukin-12 p70 secretion by lipopolysaccharide-stimulated adherent cells from the peripheral blood mononuclear cells of 27 burn and trauma patients measured by enzyme-linked immunoadsorbent assay on 53 occasions at serial intervals after injury and compared with 18 normal control subjects. The patients’ adherent cells produced significantly less interleukin-12 than cells from normal persons at multiple intervals beginning early after injury.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 2. Production of interleukin-12 (IL-12) and IL-10 by lipopolysaccharide-stimulated peripheral blood mononuclear cells (PBMCs) of burn and trauma patients at serial intervals after injury. Thirty-eight observations of IL-10 production were made in 18 patients and 21 observations of IL-12 production in 8 of the same patients. PBMC IL-12 production was significantly diminished in the 8- to 14-day interval after injury when compared with simultaneously studied normal subjects. At the same interval, IL-10 production by the patients’ PBMCs was significantly elevated.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques:

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 3. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed 10 days after injury. Burn mice receiving 5 ng interleukin-12 (IL-12) every other day for 9 days beginning on the day of injury had a survival similar to that of sham burn mice. IL-12 in a 10-ng dose appeared to have a less favorable effect on survival, and the addition of indomethacin to the 5-ng dose of IL-12 produced inferior results. IL-12 treatment did not affect the survival of sham burn mice.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 4. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed on day 10 after injury. The burn mice treated with 5 ng interleukin-12 (IL-12) every other day had a survival similar to that of sham burn controls, even though the IL-12 treatment was continued through day 11, beyond the time of cecal ligation and puncture. Untreated burn animals had the expected high death rate.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 5. Survival of groups of 12 to 15 burn and sham burn mice after cecal ligation and puncture performed on day 10 after injury. The dose of 1 ng interleukin-12 (IL-12) given every other day in burn animals and continued beyond the time of cecal ligation and puncture produced survival equivalent to that of sham burn controls.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 7. Interferon-gamma production by anti-CD3 antibody-stimulated splenocytes from groups of 5 to 10 burn and sham burn animals 10 days or 13 days after injury (after cecal ligation and puncture on day 10). The burn groups treated with interleukin-12 (IL-12) produced significantly more interferon-gamma than untreated animals. S and S/12 indicate untreated and IL-12–treated sham mice. B and B/12 indicate untreated and IL-12–treated burn mice.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 8. Tumor necrosis factor-alpha production by lipopolysaccharide-stimulated adherent splenocytes from groups of 5 to 10 burn and sham burn animals 10 days or 13 days after injury (after cecal ligation and puncture on day 10). Splenocyte production of tumor necrosis factor-alpha was markedly increased in the burn animals treated with interleukin-12.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 9. Production of interleukin-10 (IL-10) by anti-CD3–stimulated splenocytes harvested from groups of 5 to 10 burn and sham burn mice before cecal ligation and puncture on day 10 after injury or on day 13 (after cecal ligation and puncture on day 10). IL-12 treatment increased IL-10 production by burn splenocytes, both before and after cecal ligation and puncture. Splenocytes from untreated burn animals produced more IL-10 than sham burn splenocytes at both intervals.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation, Produced

Journal:

Article Title: Injury Induces Deficient Interleukin-12 Production, But Interleukin-12 Therapy After Injury Restores Resistance to Infection

doi:

Figure Lengend Snippet: Figure 10. Total mononuclear cells per spleen in groups of five burn and sham burn animals at day 10 after the burn and at day 13, 3 days after cecal ligation and puncture was performed on day 10. The mononuclear cell population in the spleen was markedly reduced 3 days after cecal ligation and puncture, and treatment with interleukin-12 significantly increased the splenic mononuclear cell population of burn animals.

Article Snippet: In the human assay, the capture antibody was a mouse antihuman IL-12 p70 monoclonal antibody (MAB611) and the detection antibody was a biotinylated goat antihuman IL-12 monoclonal antibody (BAF219) (both antibodies from R&D Systems, Minneapolis, MN).

Techniques: Ligation