Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) Immunohistochemical analyses of ERβ, MMP-1, MMP-2 and MMP-9 in NSCLC tissue. NSCLC specimens were immunostained with ERβ, MMP-1, MMP-2 and MMP-9-specific antibodies, as mentioned in ‘‘Materials and methods’’ section. During immunostaining, positive cells appeared as yellowish brown or contained yellowish brown granules (Original magnification 200×(left) and 400×(right)). (B) The proportion of lymph node metastasis in NSCLC patients with different ERβ IHC scores as described in the Materials and Methods section (-: > 4; + : 4∼8; ++ : 8∼12; +++ : 12∼16); (C) Immunohistochemical staining scores in 222 NSCLC samples expressed by box plots. Significance of mean differences in staining scores between “Metastasis” and “No Metastasis” group is shown ( *p = 0.017 < 0.05 ). (D) Significance of mean differences in staining scores between “Well/Moderate” and “Poor” tumor differentation grade group ( *p = 0.035 < 0.05 ). (E) Western blot analysis of ERβ expression in 8-pair cases of metastatic lymph node tissues and primary NSCLC tumor tissues. GAPDH was used as a control.

Article Snippet: Rabbit anti-human ERβ(1:200, Proteintech, CA, USA), mouse anti-human MMP-1(1:200, Santa Cruz Biotechnology, CA, USA), mouse anti-human MMP-2 (1:200, Santa Cruz Biotechnology, CA, USA), and rabbit anti-human MMP-9(1:500,Proteintech, CA, USA) antibodies were used for staining.

Techniques: Immunohistochemical staining, Immunostaining, Staining, Western Blot, Expressing

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: Correlation between ERβ expression with MMP-1, MMP-2 and MMP-9 in 222 cases of non-small cell lung carcinoma

Article Snippet: Rabbit anti-human ERβ(1:200, Proteintech, CA, USA), mouse anti-human MMP-1(1:200, Santa Cruz Biotechnology, CA, USA), mouse anti-human MMP-2 (1:200, Santa Cruz Biotechnology, CA, USA), and rabbit anti-human MMP-9(1:500,Proteintech, CA, USA) antibodies were used for staining.

Techniques: Expressing

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A-B) Western blotting analysis of ERα, ERβ, MMP-1, MMP-2 and MMP-9 expression in cultured NSCLC cell lines (A549 and H1793) and breast cancer cell lines (MCF-7). (C-D) The synchronized cells were treated with E2 at different concentrations (0 nM, 0.1 nM, 1 nM, 10 nM and 100 nM) for 2 days. The protein expression of ERβ and MMP-2 was analyzed using Western blot. The data represent mean ± SEM from three different experiments. E2 stimulated both ERβ and MMP-2 response in A549 cell lines in a dose-dependent manner. (E-F) The synchronized cells were treated with E2 at different time points (0 h, 1 h, 6 h, 12 h, 24 h and 48 h) at concentrations of 10 nM. The protein expression of ERβ and MMP-2 was analyzed using Western blot. The data represent mean ± SEM from three different experiments. The data represent mean ± SEM from three different experiments. E2 stimulated both ERβ and MMP-2 response in A549 cell lines in a time-dependent manner. GAPDH was used as a control.

Article Snippet: Rabbit anti-human ERβ(1:200, Proteintech, CA, USA), mouse anti-human MMP-1(1:200, Santa Cruz Biotechnology, CA, USA), mouse anti-human MMP-2 (1:200, Santa Cruz Biotechnology, CA, USA), and rabbit anti-human MMP-9(1:500,Proteintech, CA, USA) antibodies were used for staining.

Techniques: Western Blot, Expressing, Cell Culture

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) The migration in wound-healing assay of A549 cells after treatment with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1 μM) and Ful (1 μM) for 24 h (magnification 40×); (B) The effect on wound closure (percentage) in A549 cells ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ); (C) Representative images of Transwell assays to assess A549 cell invasion and migration; (D) The effect of transwell migration assays in A549 cells after treating with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1μM) and Ful(1μM) for 24 h; The effect of transwell migration assays in H1793 cells after treatment with the same drugs for 72 h; (E) Representative images of Transwell assays for assessment of H1793 cell invasion and migration; (F) The effect of transwell migration assays in H1793 cells after treatment with DMSO or E2 (10 nM), PPT (10 nM), DPN (10 nM), E2+Ful (1 μM) and Ful (1 μM) for 24 h. The effect of transwell migration assays in H1793 cells after treatment with the same drugs for 72 h; (G-H) Western blot analysis of ERβ, MMP-2, MMP-9, p-p38MAPK, p38MAPK, pAKT and tAKT protein levels at 48 h in A549 cells, respectively. GAPDH was used as a control. All data are shown as the mean ± SD. Results represent three independent experiments. Student's t-test was carried out ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ).

Article Snippet: Rabbit anti-human ERβ(1:200, Proteintech, CA, USA), mouse anti-human MMP-1(1:200, Santa Cruz Biotechnology, CA, USA), mouse anti-human MMP-2 (1:200, Santa Cruz Biotechnology, CA, USA), and rabbit anti-human MMP-9(1:500,Proteintech, CA, USA) antibodies were used for staining.

Techniques: Migration, Wound Healing Assay, Western Blot

Journal: Oncotarget

Article Title: Estrogen promotes tumor metastasis via estrogen receptor beta-mediated regulation of matrix-metalloproteinase-2 in non-small cell lung cancer

doi: 10.18632/oncotarget.16992

Figure Lengend Snippet: (A) After bilateral ovariectomy A549 cells (5 × 10 6 /100 μL) suspended in PBS were injected into 4-week-old female BALB/c nude mice via the tail vein, mice were randomly divided into five groups (N = 5/group): E2(0.09 mg/kg), PPT(1.05 mg/kg), DPN(1.89 mg/kg), E2+ Ful(1.46 mg/kg) and blank control. The lungs were removed after 4 weeks subcutaneously under drug treatment. Gross appearance of metastatic lung tumor nodes in different groups is indicated by bright yellow arrows. (B) Representative pathological image of metastatic A549 tumor at magnification 100ificatione pathological image of groups (N = 5/group): The number of metastatic nodules in the lungs (lung nodules number in every mouse of each group), metastatic index in different groups mentioned in ‘‘Materials and methods’’ section for the experimental. (C-E) The mean lung wet weight of each group, The number of metastatic nodules in the lungs (lung nodules number in every mouse of each group), metastatic index in different groups mentioned in ‘‘Materials and methods’’ section for the experimental. (F) Protein expression of ERβ, MMP-2, MMP-9, p-p38MAPK, p38MAPK, pAKT and tAKT in murine lung metastatic nodes was analyzed using Western blot, and (G) the detection of optical density. All data are expressed as the mean ± SD. Student's t-test was carried out ( *p < 0.05 vs. control group, #p < 0.05 vs. E2 group ).

Article Snippet: Rabbit anti-human ERβ(1:200, Proteintech, CA, USA), mouse anti-human MMP-1(1:200, Santa Cruz Biotechnology, CA, USA), mouse anti-human MMP-2 (1:200, Santa Cruz Biotechnology, CA, USA), and rabbit anti-human MMP-9(1:500,Proteintech, CA, USA) antibodies were used for staining.

Techniques: Injection, Expressing, Western Blot