CD9 antigen mRNA is induced by hypertonicity in two renal epithelial cell lines

1996 ◽  
Vol 270 (1) ◽  
pp. C253-C258 ◽  
Author(s):  
D. Sheikh-Hamad ◽  
J. D. Ferraris ◽  
J. Dragolovich ◽  
H. G. Preuss ◽  
M. B. Burg ◽  
...  
Keyword(s):  
Stress Response ◽  
Cell Lines ◽  
Gene Transcription ◽  
Hyperosmotic Stress ◽  
Surface Glycoprotein ◽  
Specific Gene ◽  
Organic Osmolytes ◽  
Cell Surface Glycoprotein ◽  
Hypertonic Medium ◽  
Mrna Induction

In diverse organisms, cells adapt to hyperosmotic stress by accumulating organic osmolytes. Mammalian renal medullary cells are routinely under osmotic stress. Two renal cell lines, Madin-Darby canine kidney (MDCK) and PAP-HT25, have been widely used to study mammalian osmotic regulation. In these epithelial cells, extracellular hypertonicity induces gene transcription of proteins directly involved in the metabolism and transport of organic osmolytes. This induction is relatively specific and not part of a generalized stress response. Little is known about the signal transduction pathway between cellular detection of extracellular osmolality and increased specific gene transcription. Here, using differential mRNA display polymerase chain reaction on MDCK cells in isotonic vs. hypertonic medium, we identify a cDNA product corresponding to CD9 antigen mRNA. CD9 antigen is a cell surface glycoprotein originally found in cells of the immune system. Although CD9 antigen has been structurally characterized, its function is unclear. We further demonstrate that CD9 antigen mRNA is present in MDCK and PAP-HT25 cells and that its mRNA abundance is induced by extracellular hypertonicity, but not by heat stress. Also, we show that accumulation of organic osmolytes markedly attenuates the CD9 mRNA induction, as only recently demonstrated with genes involved in the hyperosmotic stress response. This suggests a role for CD9 antigen in this response.

Distribution of cell surface glycoprotein CD9 (P24) antigen on megakaryocyte lineage leukemias and cell lines

1990 ◽  
Vol 35 (1) ◽  
pp. 65-67 ◽  
Author(s):  
Nobutaka Imamura ◽  
Deodatus M. Mtasiwa ◽  
Hiromi Ota ◽  
Tominari Inada ◽  
Atsushi Kuramoto
Keyword(s):  
Cell Surface ◽  
Cell Lines ◽  
Surface Glycoprotein ◽  
Cell Surface Glycoprotein ◽  
P24 Antigen

Kidney aldose reductase gene transcription is osmotically regulated

1992 ◽  
Vol 262 (3) ◽  
pp. C776-C782 ◽  
Author(s):  
F. L. Smardo ◽  
M. B. Burg ◽  
A. Garcia-Perez
Keyword(s):  
Gene Transcription ◽  
Aldose Reductase ◽  
Synthesis Rate ◽  
Protein Synthesis Rate ◽  
Transcription Rate ◽  
Organic Osmolytes ◽  
Animal Cells ◽  
Hypertonic Medium ◽  
Hypertonic Stress ◽  
Reductase Mrna

Cells generally adapt to long-term hypertonic stress by accumulating organic osmolytes. PAP-HT25 renal medullary cells in hypertonic medium accumulate sorbitol through a reaction catalyzed by aldose reductase and betaine through osmotically regulated transport. Hypertonicity increases aldose reductase protein synthesis rate by elevating its mRNA abundance. To test whether the rise in aldose reductase mRNA is due to enhanced transcription, PAP-HT25 cells adapted to isotonic medium were switched to hypertonic medium, and transcription rate was measured by nuclear run-on. Aldose reductase transcription rate peaked at 17-fold the isotonic level after 12 h of hypertonicity. Then, transcription fell as sorbitol and betaine accumulated. Transcription stabilized at fivefold the isotonic level within days. Aldose reductase mRNA stability was not significantly different between the hypertonic and isotonic steady states. Thus aldose reductase mRNA is osmotically regulated through changes in its transcription. The osmotically induced rise in aldose reductase transcription is blunted by the accumulation of intracellular betaine and is exaggerated and prolonged by preventing the accumulation of both sorbitol (by aldose reductase inhibition) and betaine (by removal from the medium). This study presents the first description of osmoregulated gene transcription in animal cells.

Exploring the potential of mucin 13 (MUC13) as a biomarker for carcinomas and other diseases

2018 ◽  
Vol 56 (11) ◽  
pp. 1945-1953 ◽  
Author(s):  
Panagiota S. Filippou ◽  
Annie H. Ren ◽  
Dimitrios Korbakis ◽  
Lampros Dimitrakopoulos ◽  
Antoninus Soosaipillai ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Surface Glycoprotein ◽  
Serum Samples ◽  
Cell Surface Glycoprotein ◽  
Ovarian Cancer Cell Lines ◽  
Human Sera ◽  
A Cell ◽  
Inflammatory Bowel

Abstract Background: Mucin 13 (MUC13) is a cell surface glycoprotein aberrantly expressed in a variety of epithelial carcinomas. Thus far, the role of MUC13 in various diseases remains elusive. To the best of our knowledge, this is the first study to examine the potential of MUC13 as a serum biomarker in a variety of carcinomas and other conditions. Methods: We developed a recombinant MUC13 protein, mouse monoclonal antibodies and enzyme immunoassay (ELISA) for MUC13. We used this assay to measure MUC13 levels in the supernatants of cancer cell lines and a large cohort of serum samples from healthy and diseased individuals. Results: MUC13 is secreted from cancer cell lines, with highest levels found in ovarian cancer cell lines. MUC13 levels in human sera were significantly increased in patients with renal failure and 20%–30% of patients with ovarian, liver, lung and other cancers. MUC13 was also elevated in 70% of patients with active cutaneous melanoma, but not uveal melanoma. Furthermore, we identified significant MUC13 elevations in the serum of patients with vasculitis (ANCA-positive) autoantibodies, but not in those with inflammatory bowel disease. Conclusions: Serum MUC13 is frequently elevated not only in a variety of malignant cases but also in some benign pathologies, thus appearing to be a non-specific disease biomarker. Nonetheless, serum MUC13 is clearly highly elevated in some carcinoma patients, and its relationship with tumor progression in this context warrant further research. Future studies that examine the correlation between serum MUC13 levels to stage of cancer could elucidate prognostic potential.

scholarly journals Reactivation of apolipoprotein II gene transcription by cycloheximide reveals two steps in the deactivation of estrogen receptor-mediated transcription.

1994 ◽  
Vol 14 (3) ◽  
pp. 1733-1742 ◽  
Author(s):  
M G Sensel ◽  
R Binder ◽  
C B Lazier ◽  
D L Williams
Keyword(s):  
Gene Expression ◽  
Estrogen Receptor ◽  
Protein Synthesis ◽  
Cell Lines ◽  
Gene Transcription ◽  
Mrna Stability ◽  
Stable Expression ◽  
Chicken Liver ◽  
Mrna Induction ◽  
Estrogen Withdrawal

In this report, we describe apolipoprotein II (apoII) gene expression in cell lines derived by stable expression of the chicken estrogen receptor in LMH chicken hepatoma cells. In cell lines expressing high levels of receptor (LMH/2A), apoII gene expression is increased by estrogen 300-fold compared with levels in the receptor-deficient parent LMH line. LMH/2A cells show apoII mRNA induction and turnover kinetics similar to those in chicken liver. Inhibition of protein synthesis with cycloheximide (CHX) or puromycin following estrogen withdrawal superinduces apoII mRNA without affecting apoII mRNA stability. Superinduction is due to an estrogen-independent reactivation of apoII gene transcription. The apoII gene can be reactivated by CHX for up to 24 h following hormone withdrawal, suggesting that the gene is in a repressed yet transcriptionally competent state. These results reveal two distinct events necessary for termination of estrogen receptor-mediated transcription. The first event, removal of hormone, is sufficient to stop transcription when translation is ongoing. The second event is revealed by the CHX-induced superinduction of apoII mRNA following hormone withdrawal. This superinduction suggests that deactivation of estrogen receptor-mediated transcription requires a labile protein. Furthermore, reactivation of apoII gene expression by CHX and estrogen is additive, suggesting that estrogen is unable to overcome repression completely. Thus, a labile protein may act to repress estrogen receptor-mediated transcription of the apoII gene.

Mesothelin expression in patients as a novel target in gastric cancer.

2014 ◽  
Vol 32 (3_suppl) ◽  
pp. 61-61 ◽  
Author(s):  
Ronan Joseph Kelly ◽  
Ira Pastan ◽  
Morgan L Cowan ◽  
Elizabeth Montgomery ◽  
Raffit Hassan ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cell Lines ◽  
Cancer Cell ◽  
Gastric Cancer Cell ◽  
Stage Iv ◽  
Cancer Cell Lines ◽  
Surface Glycoprotein ◽  
Cell Surface Glycoprotein ◽  
Gastric Cancer Cell Lines ◽  
Mesothelin Expression

61 Background: Novel targets and therapeutics are of profound importance if we are to improve outcomes in gastric cancer. This study assessed the incidence of mesothelin expression in tumors of Western patients with gastric cancer. In addition, gastric cancer cell lines were then tested for their sensitivity to SS1(dsFv)PE38 (SS1P), a mesothelin-targeted immunotoxin. Methods: Tissue specimens from 127 patients with gastric or gastro-esophageal junction (GEJ) tumors were examined by immuno-histochemistry (IHC) for intensity of staining for mesothelin. Expression of HER2-neu, e-cadherin and c-Met were also assessed. Tumors were considered positive for mesothelin if moderate cytoplasmic/membranous or luminal staining was present in at least 10% of the cells. Gastric cancer cell lines were assessed for surface mesothelin expression by flow cytometry and the viability of cells treated with SS1P was measured. Results: Gastric and GEJ cancers were mesothelin positive in 64/127 tumors (50%) while only 9 tumors (7%) were Her2-neu IHC +3 positive and 8 tumors (6%) were c-Met positive (c-Met low 5% and c-Met high 1%). Mesothelin expression rates changed from stage I tumors to stage IV tumors (39% to 56% respectively). We found that both the AGS and MKN28 gastric cancer cell lines express mesothelin and are sensitive to the immunotoxin with EC50 values in the low picomolar range (0.4 and 0.3 ng/mL, respectively). Conclusions: Mesothelin is a cell surface glycoprotein that is expressed in 50% of resected gastric cancers. Gastric cancer cell lines were exquisitely sensitive to SS1P. The high expression of mesothelin and its sensitivity to the immunotoxin SS1P makes it an attractive tumor associated antigen for therapeutic targeting. Based on these observations clinical trials involving novel mesothelin targeted drugs in gastric cancer are currently in development.

scholarly journals CD133 Expression and Identification of CD133/nestin Positive Cells in Rhabdomyosarcomas and Rhabdomyosarcoma Cell Lines

2011 ◽  
Vol 34 (6) ◽  
pp. 303-318 ◽  
Author(s):  
Jiri Sana ◽  
Iva Zambo ◽  
Jan Skoda ◽  
Jakub Neradil ◽  
Petr Chlapek ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Lines ◽  
Tumor Tissue ◽  
Surface Glycoprotein ◽  
Intermediate Filament Protein ◽  
Tissue Samples ◽  
Cell Surface Glycoprotein ◽  
Cd133 Expression ◽  
Functional Assays ◽  
Rhabdomyosarcoma Cell

Background: Co-expression of CD133, cell surface glycoprotein, and nestin, an intermediate filament protein, was determined to be a marker of neural stem cells and of cancer stem cells in neurogenic tumors.Methods: We examined the expression of CD133 and nestin in ten tumor tissue samples taken from patients with rhabdomyosarcomas and in five rhabdomyosarcoma cell lines. Immunohistochemistry and immunofluorescence were used to examine FFPE tumor tissue samples. Cell lines were analyzed by immunofluorescence, immunoblotting, flow cytometry, and RT-PCR. Functional assays (clonogenicin vitroassay and tumorigenicin vivoassay) were also performed using these cell lines.Results: CD133 and nestin were detected in all 10 tumor tissue samples and in all 5 cell lines; however, the frequency of CD133+, Nes+, and CD133+/Nes+ cells, as well as the intensity of fluorescence varied in individual samples or cell lines. The expression of CD133 and nestin was subsequently confirmed in all cell lines by immunoblotting. Furthermore, we observed an increasing expression of CD133 in relation to the cultivation. All cell lines were positive for Oct3/4 and nucleostemin; NSTS-11 cells were also able to form xenograft tumors in mice.Conclusion: Our results represent the first evidence of CD133 expression in rhabdomyosarcoma tissue and in rhabdomyosarcoma cell lines. In addition, the co-expression of CD133 and nestin as well as results of the functional assays suggest a possible presence of cancer cells with a stem-like phenotype in these tumors.

scholarly journals Quantitation of a transformation-sensitive, adhesive cell surface glycoprotein. Decrease of several untransformed permanent cell lines.

1977 ◽  
Vol 74 (2) ◽  
pp. 649-654 ◽  
Author(s):  
K M Yamada ◽  
S S Yamada ◽  
I Pastan
Keyword(s):  
Cell Surface ◽  
Cell Lines ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cellular Transformation ◽  
Cell Types ◽  
Surface Glycoprotein ◽  
Immunofluorescent Staining ◽  
Early Passage ◽  
Cell Surface Glycoprotein ◽  
Permanent Cell

We have quantitated the transformation-sensitive, cell surface LETS glycoprotein on many untransformed cell types. By SDS-polyacrylamide gel electrophoresis, this trypsin-sensitive iodinatable glycoprotein comprises 1-3% of total cellular protein of the seven early passage cell types tested. In contrast, it constitutes less than 0.15% of the protein in four of six continuous cell lines. This decrease is reflected in alterations both in [14C]glucosamine labeling and in the immunofluorescent staining of early passage vs. these four permanent cell lines. These results help to clarify previous experiments in which CSP, a purified LETS protein, partially restored a fibroblastic phenotype to cells transformed by tumor viruses. These findings also indicate that a major decrease in this cell surface glycoprotein can occur in the establishment of a continuous cell line without resulting in cellular transformation.

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