Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: High expression of SERPING1 indicates better patient survival. (A) The two datasets, the Roessler Liver microarray and Huh7 sorafenib resistance array datasets, were analyzed and intersected to find potential candidates. SERPING1 emerged as a potential candidate and was selected for further study. (B) Immunohistochemistry staining revealed that SERPING1 expression was lower in clinical HCC tissues compared to normal tissues. The immunohistochemistry staining scores were quantified (C‐F). SERPING1 levels were significantly lower in patients with high alpha‐fetoprotein (AFP) levels, high predicted metastasis risk signature score, large tumor sizes, and advanced pathological stages. (G, H) Kaplan‐Meier survival curves of two groups of patients defined by different SERPING1 expression levels were established on the basis of Roessler liver microarray scores ( n for each group = 121, cutoff point = 10.82 (the median), log‐rank p < 0.05). Patients with high SERPING1 levels have better survival than those with low levels.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Expressing, Microarray, Immunohistochemistry, Staining

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: Association of SERPING1 level (Roessler liver array) with clinicopathologic indicators of hepatocellular carcinoma.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Cross-linking Immunoprecipitation

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: Prognostic significance of clinicopathologic indicators and SERPING1 for overall survival in the Roessler liver array.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques:

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: Sorafenib increases SERPING1 expression and cell viability. (A‐C) HCC cell viability was determined by MTT assay at 24 and 48 h after sorafenib (0–10 μM) treatment in HepG2 (A), Huh7 (B), and J7 (C) cells. Sorafenib decreased cell viability in a dose‐ and time‐dependent manner. (D–F) The SERPING1 levels in both the cell lysate (D, E) and conditioned medium (CM) (F) with various doses (5–10 μM) of sorafenib stimulation were determined using Western blotting in HepG2 (D, F) and Huh7 (E) cells. Sorafenib induced SERPING1 expression in both cell lysate and CM in HCC cells. Coomassie Brilliant Blue staining was used as a loading control.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Expressing, MTT Assay, Western Blot, Staining, Control

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: Sorafenib‐resistant cells have a higher viability rate after sorafenib treatment than parental cells. (A, B) Sorafenib‐resistant (SR) cells were established, and the cell viability of SR cells was not significantly influenced by 5 μM sorafenib at either 24 or 48 h. However, the cell viability of parental cells (PCs) was significantly decreased at 24 and 48 h after 5 μM sorafenib treatment in HepG2 (A) and Huh7 (B) cells. (C, D) The SERPING1 level was examined in the cell lysate of sorafenib‐resistant (SR; 1,2: Two resistant cell lines) HCC cells, and SERPING1 expression was induced with 5 μM sorafenib stimulation in HepG2 (C) and Huh7 (D) cells. (E) The SERPING1 levels were determined in HepG2‐PC and HepG2‐SR cells by Western blotting. SERPING1 was lower expressed in SR compared to PC.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: SERPING1 decreases cell migration. (A‐H) Huh7 (A, B, E, F) and J7 (C, D, G, H) cells were stimulated with 10 or 100 ng/mL recombinant SERPING1 protein and subjected to wound healing (A, C) and a Transwell (E, G) analyses with quantification (B, D, F, H). Treatment with recombinant SERPING1 decreased cell migration.

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Migration, Recombinant

Journal: Environmental Toxicology

Article Title: SERPING1 Reduces Cell Migration via ERK ‐ MMP2 ‐ MMP ‐9 Cascade in Sorafenib‐ Resistant Hepatocellular Carcinoma

doi: 10.1002/tox.24434

Figure Lengend Snippet: SERPING1 reverses sorafenib‐resistant cell‐induced cell migration. (A‐D) Baseline cell migration ability was higher in SR cells than in PC cells. Treatment with recombinant SERPING1 protein reduced the number of migratory cells in both PC and SR cells, as demonstrated by the Transwell (A) and wound healing (C) assays. Notably, even with SERPING1 stimulation, the number of migratory SR cells remained higher than that of PC cells. The results were also quantified (B, D).

Article Snippet: Total cell extracts (10 μg of protein) were resolved by 10% SDS–PAGE, transferred onto an Immobilon polyvinylidene difluoride membrane (Amersham Biosciences), and immunoblotted with specific primary antibodies against SERPING1 (GeneTex), p‐ERK (ABclonal, Wobum, MA, USA), ERK (ABclonal), and β‐actin (GeneTex) overnight at 4°C.

Techniques: Migration, Recombinant