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: Oncotarget

Article Title: Nicotinic acetylcholine receptors induce c-Kit ligand/Stem Cell Factor and promote stemness in an ARRB1/ β-arrestin-1 dependent manner in NSCLC

doi:

Figure Lengend Snippet: Microarray was performed in ARRB1 depleted and nicotine stimulated A549 cells Nicotine induced and ARRB1 dependent genes from the microarray data were analyzed. We identified differentially regulated genes that were regulated by nicotine in a β-arrestin-1 dependent fashion and top 10 up/down regulated genes from the list were used for prognosis prediction. Assessment of the expression of these genes for smoking revealed that SCF (highlighted in red) strongly differentiated smokers from non-smokers implying an important role of this gene in lung carcinogenesis induced by smoking

Article Snippet: A human lung cancer tissue microarray slide (Catalog number IMH-358; Imgenex, San Diego, CA) was immunostained for SCF.

Techniques: Microarray, Expressing, Control

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

Journal: bioRxiv

Article Title: Antagonizing the serotonin receptor HTR2B drives antigen-specific cytotoxic T-cell responses and controls colorectal cancer growth

doi: 10.1101/2025.02.26.640476

Figure Lengend Snippet: TCGA-COAD data were analyzed. (A) Kaplan-Meier survival analysis of HTR 2B expression (derived from TCGA data) in 220 colon adenocarcinomas patient’s cohort. “HTR 2B -high” versus “HTR 2B -low” data were segregated based on the cohort’s top or bottom 25% with intratumoral HTR 2B expression. Mantel-cox log rank test. (B) Representative images of HTR 2B and Ki67 in the control (lamina propria) and lamina propria of colon adenocarcinoma (COAD) tissue. The original magnification is 200X, and the inset images are 600X. (C) Representative images of serotonin (5-HT), neurofilament-A (NF-A) in the control (lamina propria) and lamina propria of colon adenocarcinoma (COAD) tissue IHC. Original magnification 200X. (D) RNA-seq datasets (n=30 samples) were segregated into “HTR 2B -high” and “HTR 2B -low” (highest fifteen and lowest fifteen intra-tumoral HTR 2B expression), and the differential expression of selected genes was analyzed. (E) Principal component analysis (PCA) of “HTR 2B -high” versus “HTR 2B -low” datasets (n=15 patients/group). (F) Venn diagram showing numbers of mutually exclusive or shared genes in each group. (G) A volcano plot shows differentially expressed genes (DEGs) of “HTR 2B -high” versus “HTR 2B -low” datasets (n=15 patients/groups) are shown. (H) Heatmap statistics differentially expressed selected gene sets. (I) Gene ontology of altered biological processes. (J) Gene set enrichment analysis (GSEA) of the curated gene sets that affect the regulatory and effector functions of the T cells. (K) Intra-tumoral immune cell signatures were analyzed by deconvolution using TIMER2.0 of HTR 2B -high and -low datasets. (L) A TIDE computational analysis of the association between the tumor-infiltrating CD8 T cells (CTL) level (expression of CD8A, CD8B, GZMA, GZMB, and PRF1. Overall, patient survival with the intratumoral HTR 2B gene expression level. For each patient cohort, tumor samples were divided into HTR 2B -high (samples with HTR 2B expression one standard deviation above the average, shown in the left survival plot) and HTR 2B -low (remaining samples shown in the right survival plot) groups.

Article Snippet: The human colon adenocarcinoma tissue microarray (Novus Biological, Catalog # NBP2-78088) was stained with the antibodies and detected using chromogenic reagent DAB.

Techniques: Expressing, Derivative Assay, Control, RNA Sequencing, Gene Expression, Standard Deviation

Journal: Clinical 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. 1. Expression of CLIC4 is altered in several human cancer types. A, tumor array representing cDNAs derived from the ‘‘matched’’normal (N) and tumor (T) tissues (paired horizontally) of multiple human cancer types is probed with radioactively labeled CLIC4 or ubiquitin (inset) DNA probes. B, oncomine database (University of Michigan Comprehensive Cancer Center) was searched with‘‘CLIC4’’ as a query.Two examples of statistically significant changes (prostate/Lapointe/ Stanford University/P = 1.8 1021and lung/Bhattacharjee/Harvard Medical School/P = 3 109). Blue columns, normal tissues; red columns, cancerous tissues. Brackets,1.5 times interquartile range; dots, outliers; line within box, median; box boundaries, upper and lower quartiles.Two independent studies. C, tumor lysate dot blot representing 200 tumor samples (600 ng per spot) is probed with NH2-terminal CLIC4 antibody. ‘‘Matched’’normal (solid gray arrowhead) and tumor (solid black arrowhead) tissue samples are spotted vertically as a pair. Open circles are marker spots for an orientation.The experiment was repeated twice, and independent experiments with 10-fold less amount of lysate loaded per spot showed similar results.

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, Derivative Assay, Labeling, Ubiquitin Proteomics, Dot Blot, Marker

Journal: Clinical 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