Journal:

Article Title: Increased RNA-Induced Silencing Complex (RISC) Activity Contributes to Hepatocellular Carcinoma

doi: 10.1002/hep.24216

Figure Lengend Snippet: Inhibition of SND1 enzymatic activity inhibits cell viability and AEG-1 function. A. Hep-pc-4 and Hep-AEG-1-14 cells were treated or not with 50, 100 and 200 μM pdTp for 1 to 4 days and cell viability was measured by standard MTT assay. B. Hep-pc-4, Hep-AEG-1-14, Hep3B and QGY-7703 cells were treated with 100 μM pdTp and colony formation assay was performed. The colonies were scored after two weeks. Data represent mean± SEM of three independent experiments. C. Analysis of SND1 expression in tissue microarray by immunohistochemistry. ˆ: p<0.05, #: p<0.02 and *: p<0.01 vs the corresponding data point in Hep-pc-4 cells.

Article Snippet: Tissue microarray and immunostaining Human HCC tissue microarrays were obtained from Imgenex Corp. Two tissue microarrays were used: one containing 40 primary HCC, 10 metastatic HCC and 9 normal adjacent liver samples (Imgenex; IMH-360), the other containing 46 primary HCC and 13 metastatic HCC (Imgenex; IMH-318).

Techniques: Inhibition, Activity Assay, MTT Assay, Colony Assay, Expressing, Microarray, Immunohistochemistry

Journal:

Article Title: Increased RNA-Induced Silencing Complex (RISC) Activity Contributes to Hepatocellular Carcinoma

doi: 10.1002/hep.24216

Figure Lengend Snippet: Immunoperoxidase staining of normal liver and different stages of HCC by tissue microarray using anti-SND1 antibody

Article Snippet: Tissue microarray and immunostaining Human HCC tissue microarrays were obtained from Imgenex Corp. Two tissue microarrays were used: one containing 40 primary HCC, 10 metastatic HCC and 9 normal adjacent liver samples (Imgenex; IMH-360), the other containing 46 primary HCC and 13 metastatic HCC (Imgenex; IMH-318).

Techniques: Immunoperoxidase Staining, Microarray, Staining

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Representative immunofluorescent staining of cultured mesothelioma cells using AX10 antibody (a), immunohistochemical staining (b), and secondary antibody‐drug conjugate assay in vitro (c). (a) AX10 immunoreactivity in MPM‐1, −2, and −3 cells, representing sarcomatoid, epithelioid, and biphasic type mesothelioma, respectively. All MPM‐1, −2, and −3 cells exhibited AX10 antibody immunoreactivity at the cell surface. The staining was analyzed using a Guava easyCyte cell analyzer and accompanying software to obtain a one‐parameter log histogram. (b) AX10 immunoreactivity in various mesothelioma tissue specimens. Weak or no AX10 immunoreactivity was detected in five out of 10 epithelioid mesothelioma tissues (a). One out of five biphasic mesotheliomas exhibited AX10 immunoreactivity in spindle sarcomatoid components (arrow) but weak immunoreactivity in epithelioid components (arrowhead) (b). Five out of six sarcomatoid mesothelioma tissues exhibited strong AX10 immunoreactivity (c). Little AX10 immunoreactivity was detected in normal human tissues. No significant AX10 immunoreactivity was detected in the lung (d) (pleural mesothelial cells; insert) tissue specimens. Weak AX10 immunoreactivity was detected in myofibrous cells in the uterus (e). We did not detect any significant AX10 immunoreactivity in the brain, liver, or kidney, whereas strong AX10 immunoreactivity was observed in a nonmelanocytic (hypomelanocytic) melanoma tissue sample that was supplementally included in the microarray (f) (staining without AX10 antibody; insert). (c) MPM‐1 sarcomatoid mesothelioma cells were incubated with AX10 at 10, 100, and 1000 ng/mL followed by incubation with anti‐murine IgG (Fc) antibody conjugated to duocarmycin. Representative staining with Annexin V‐PI is presented. Note the dose‐dependent Annexin V‐positive and PI‐negative apoptotic MPM‐1 cells in the presence of AX10 antibody

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Staining, Cell Culture, Immunohistochemical staining, In Vitro, Software, Microarray, Incubation

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: AX10 does not affect cell proliferation, but significantly decreases Matrigel invasion activity of MPM‐1 sarcomatoid mesothelioma cells in vitro. (a) Representative cell proliferation assay. At 24 h, the cell number was 1.80 ± 0.10 (mock) and 1.77 ± 0.06 (AX10). Respective numbers at 48 h were 2.40 ± 0.10 (mock) and 2.37 ± 0.12 (AX10), while at 72 h they were 3.90 ± 0.20 (mock) and 4.20 ± 0.61 (AX10). The data represent means ± SD from triplicate assays (Student's t ‐test, p > 0.5). (b) AX10 significantly reduced Matrigel invasion activity of MPM‐1 cells (Student's t ‐test, p < 0.01). The number of invading cells was 59.7 ± 7.02 (mock) and 10.3 ± 1.52 (AX10) at 24 h, and 210.7 ± 11.4 (mock) and 15.0 ± 3.00 (AX10) at 48 h. Data from triplicate assays are expressed as means ± SD ( n = 3). (c) Cells that migrated to the lower surface of the membrane are shown (48 h). Original magnification, ×100

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Activity Assay, In Vitro, Proliferation Assay, Membrane

Journal: Thoracic Cancer

Article Title: Tumor suppressor effect of an antibody on xenotransplanted sarcomatoid mesothelioma cells

doi: 10.1111/1759-7714.14591

Figure Lengend Snippet: Inhibitory effect of AX10 on MPM‐1 xenotransplanted sarcomatoid mesothelioma cell proliferation. (a) Inoculation of AX10 antibody delayed the growth of xenotransplanted MPM‐1 sarcomatoid mesothelioma tumors. On day 0, SCID‐NOD mice were subcutaneously implanted with MPM‐1 cells. The following day, day 3, the mice were administered AX10 antibody or vehicle only by intraperitoneal injection and weekly thereafter as indicated by arrows. Values are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test (* p < 0.01). (b) On day 42, the xenotransplanted tumors were excised to determine their weight. Total tumor weights are represented as means ± standard error for n = 5 mice. Statistical significance was measured by a two‐sided unpaired Student's t ‐test ( p < 0.01). (c) Gross and histological appearance of a representative xenotransplanted tumor. Arrowhead indicates the tumor without AX10 antibody, while the arrow indicates the small tumor remaining following weekly AX10 injection. Note the elimination of tumor cells, which were histologically replaced by regenerative muscle in mice inoculated with AX10 antibody. Scale bar indicates 100 μm

Article Snippet: Tissue microarrays composed of mesothelioma (Cat. No. MS801b) and Food and Drug Administration (FDA) normal organ tissue arrays (Cat. No. NBP2‐78057) were purchased from US Biomax and Novus Biologicals, respectively.

Techniques: Injection

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Reciprocal modifications of CLIC4 in tumor epithelium and stroma mark malignant progression of multiple human cancers.

doi: 10.1158/1078-0432.CCR-06-1562

Figure Lengend Snippet: Fig. 2. CLIC4 expression and subcellular localization are altered in human tumors. Human normal and matched tumor (roman numerals, tumor stage) tissue sections representing (A) esophagus, (B) kidney, and (C) colon from the tissue microarrays are immunostained with anti-CLIC4 antibody and visualized by bright-field microscopy under 40 magnification. Inset, lower magnification (10). Black arrows, CLIC4 localized to the nucleus in the normal tissues. Similar staining patterns of CLIC4 were found in multiple human tumor types. Representative immunohistochemical stainings.

Article Snippet: Immunohistochemistry, immunofluorescence, and tissue microarray staining Human tissue microarrays were obtained from multiple sources: TARP tissue arrays (National Cancer Institute), ‘‘matched’’ human tumor tissue arrays (Imgenex, San Diego, CA), Food and Drug Administration–standard normal and tumor tissue arrays (Biochain, Hayward, CA) and human cancer screen tissue arrays (Clinomics), prostate tumor arrays (Baylor Specialized Programs of Research Excellence), and human tissue arrays (Accumax) were used for CLIC4 expression and localization studies.

Techniques: Expressing, Microscopy, Staining, Immunohistochemical staining