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: Oncogenesis

Article Title: Chondroitin sulfate synthase 1 enhances proliferation of glioblastoma by modulating PDGFRA stability

doi: 10.1038/s41389-020-0197-0

Figure Lengend Snippet: a Comparison of CHSY1 gene expression in glioma subtypes and normal brain tissue in the REMBRANDT glioma microarray database. ** P < 0.01, **** P < 0.0001. b High expression of CHSY1 was associated with worse overall survival in glioma patients. The high and low expression groups were divided by median expression level of CHSY1 in 329 cases. These data were from the REMBRANDT database ( http://www.betastasis.com/glioma/rembrandt/ ). c Immunohistochemistry of CHSY1 (upper panel) and CS56 (lower panel) on tissue array contains 85 primary glioma cases. The staining was visualized in brown color with a 3,3-diaminobenzidine liquid substrate system. All sections were counterstained with hematoxylin. Representative images of four glioma cases with different staining intensities are shown. Amplified images are shown at the bottom right of each image. Scale bars, 50 μm. Arrows indicate positive stained glioma cells. d Representative images of CHSY1 staining on normal brain tissue ( n = 5). e Statistical analysis of immunohistochemistry in glioma tissue array. Mann–Whitney U -test was used. P -values are shown at top. f Expression of CHSY1 in glioma cell lines and normal human brain tissue. The protein expression was analyzed by western blotting. Total loading protein is shown at bottom. Relative expression levels to total brain tissue form three independent blots are shown at the right.

Article Snippet: Adult normal human brain tissue lysates were purchased from Novus Biologicals.

Techniques: Comparison, Gene Expression, Microarray, Expressing, Immunohistochemistry, Staining, Amplification, MANN-WHITNEY, Western Blot

Journal: Cell reports

Article Title: Deregulation of STING Signaling in Colorectal Carcinoma Constrains DNA-Damage Responses and Correlates With Tumorigenesis

doi: 10.1016/j.celrep.2015.12.029

Figure Lengend Snippet: (A) Immunoblot of STING in hTERT fibroblasts, normal human colon epithelial (FHC) and a series of human colon cancer cell lines. (B) ELISA analysis of human Interferon β production in the media of cells (same as A) transfected with 3μg/ml polyIC or dsDNA90 or mock transfected for 16 hours. (C) qPCR analysis of human CXCL10 expression in cells (same as A) transfected with 3μg/ml dsDNA90 or mock transfected for 3 hours. (D) qPCR analysis of human IL1B expression in cells same as C. Data is representative of at least two independent experiments. Error bars indicate s.d. *, p<0.05; **, p<0.01; ***, p<0.001; Student’s t-test. (E) Microarray analysis of gene expression in indicated normal and colon cancer cells mock transfected or transfected with 3μg/ml dsDNA90 for 3 hours. Highest variable genes are shown. Rows represent individual genes; columns represent individual samples. Pseudo-colors indicate transcript levels below (green), equal to (black), or above (red) the mean. Scale represents the intensity of gene expression (log10 scale ranges between −3 and 3). (F) Fold change values of highest variable genes shown in E. See also Figure S1 and S2.

Article Snippet: Immunohistochemistry and Histological Analysis Tissue Microarray was purchased from Origene.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Expressing, Microarray, Gene Expression

Journal: Cancers

Article Title: The Peptide-Drug Conjugate TH1902: A New Sortilin Receptor-Mediated Cancer Therapeutic against Ovarian and Endometrial Cancers

doi: 10.3390/cancers14081877

Figure Lengend Snippet: Expression of SORT1 in ovarian healthy and tumoral tissues. Immunohistochemical staining of SORT1 in tissue microarrays (TMAs) was used to measure the level of SORT1 expression in healthy tissues and in ovarian tumors. With the immunohistochemical (IHC) scoring system used, the level of expressed SORT1 ranged from 0 to a maximum of 12. ( A ) Representative SORT1 staining by IHC in ovarian healthy, primary and metastases biopsies. Nuclei are stained blue with hematoxylin. Arrows point to ovarian healthy epithelia. Black bars represent 50 µm. ( B ) IHC scores from healthy tissues ( n = 20), benign tumors ( n = 18), malignant tumors ( n = 45) and metastases ( n = 10). ( C ) RT-qPCR was used to quantify the transcript levels of SORT1 in healthy and cancerous ovarian tissues. The SORT1 transcript levels in tumor samples were segregated according to tumor grade and then compared to the values from healthy ovarian tissues. n = 8 for healthy and for Grade I tumor samples, n = 9 for Grade II, n = 17 for Grade III and n = 7 for Grade IV tumor samples. ( D ) The mean IHC scores for SORT1 is shown for healthy ovarian tissue ( n = 20), benign tumors ( n = 18), LGSC ( n = 21), HGSC ( n = 6), CCC ( n = 5), MC ( n = 3), EC ( n = 13), TCC ( n = 5) and GC and NE ( n = 20). All scatter plots include lines showing the means ± SEM, and where each point represents an individual tissue sample.

Article Snippet: The tissue microarrays (#NBP2-30170 for normal tissues, #NBP2-30290 for ovarian cancers and #NBP2-30330 for diverse cancers) were purchased from Novus Biologicals (Toronto, ON, Canada).

Techniques: Expressing, Immunohistochemical staining, Staining, Quantitative RT-PCR

Journal: Cancers

Article Title: The Peptide-Drug Conjugate TH1902: A New Sortilin Receptor-Mediated Cancer Therapeutic against Ovarian and Endometrial Cancers

doi: 10.3390/cancers14081877

Figure Lengend Snippet: Expression of SORT1 in endometrial healthy tissues and endometrial tumor biopsies. Immunohistochemical staining of SORT1 in tissue microarrays (TMAs) was used to measure the level of SORT1 expression in healthy tissues and in endometrial tumors. ( A ) Representative SORT1 staining by IHC in endometrial healthy tissue and endometrial tumor biopsies. Nuclei are stained blue with hematoxylin. Black bars represent 50 µm. ( B ) SORT1 IHC scores between healthy endometrial tissues ( n = 2) and endometrial cancerous tissues ( n = 12). Scatter plot include lines showing the means ± SEM, and where each point represents an individual tissue sample.

Article Snippet: The tissue microarrays (#NBP2-30170 for normal tissues, #NBP2-30290 for ovarian cancers and #NBP2-30330 for diverse cancers) were purchased from Novus Biologicals (Toronto, ON, Canada).

Techniques: Expressing, Immunohistochemical staining, Staining