The nuclear 3,5,3'-triiodothyronine receptor in human leukaemic cell lines

1984 ◽  
Vol 105 (3) ◽  
pp. 429-432 ◽  
Author(s):  
Juan Bernal ◽  
Leif C. Andersson
Keyword(s):  
Growth Hormone ◽  
Cell Line ◽  
Cell Lines ◽  
Rat Liver ◽  
Association Constant ◽  
Erythroid Differentiation ◽  
Leukaemic Cell ◽  
Cells In Culture ◽  
Gh Cells ◽  
High Concentrations

Abstract. The 3,5,3'-triiodothyronine (T3) receptor has been studied in a series of continuously growing human leukaemic cell lines. High concentrations of receptor were found in the erythroblastoid cell line K-562. T3 was bound to the nuclei of these cells with an association constant of 3.4 × 109 m−1, and capacity 104 fmol/100 μg DNA, or 8700 molecules/nucleus. This capacity is comparable to that of rat liver or growth hormone producing cells (GH cells) in culture, and suggests that the K-562 cell line could be a useful model for the study of T3 action on erythroid differentiation.

scholarly journals Expression and function of the ghrelin axis, including a novel preproghrelin isoform, in human breast cancer tissues and cell lines

2005 ◽  
Vol 12 (4) ◽  
pp. 839-850 ◽  
Author(s):  
P L Jeffery ◽  
R E Murray ◽  
A H Yeh ◽  
J F McNamara ◽  
R P Duncan ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Growth Hormone ◽  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Lines ◽  
Growth Hormone Secretagogue ◽  
Cancer Tissues

While oestrogen, progesterone and growth factors, including growth hormone (GH), are clearly implicated in the pathogenesis of breast cancer, there is now evidence that the newly described ghrelin axis is also involved. The aims of this study were to investigate the expression of the ghrelin axis in breast cancer tissues and cell lines and to examine the effect of ghrelin on breast cancer cell proliferation in vitro. Ghrelin and its functional receptor, the growth hormone secretagogue receptor (GHSR) type 1a, were expressed in normal breast tissue and breast cancer specimens and cell lines. In contrast, the truncated GHSR type 1b isoform was exclusively expressed in breast carcinoma, suggesting that it has potential as a diagnostic marker. Ghrelin treatment significantly increases the proliferation of the MDA-MB-435 and MDA-MB-231 breast cancer cell lines in vitro. In addition, we have described the expression of a human preproghrelin isoform, exon 3-deleted preproghrelin, which encodes mature ghrelin plus a novel C-terminal peptide. Quantitative RT-PCR was used to demonstrate that this mRNA isoform is highly expressed in the MDA-MB-435 metastatic breast cancer cell line relative to the benign MCF-10A breast epithelial cell line. The unique C-terminal peptide of exon 3-deleted preproghrelin is expressed in the glandular epithelium of breast cancer tissues, with high-grade carcinoma exhibiting the strongest immunoreactivity. The data presented here suggest that components of the ghrelin axis may represent novel markers for breast cancer and potential therapeutic targets.

Survey of osmolytes in renal cell lines

1988 ◽  
Vol 255 (2) ◽  
pp. C181-C191 ◽  
Author(s):  
T. Nakanishi ◽  
R. S. Balaban ◽  
M. B. Burg
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Renal Cell ◽  
Extracellular Fluid ◽  
Organic Osmolytes ◽  
Osmolyte Accumulation ◽  
High Concentrations ◽  
Intracellular Milieu ◽  
Renal Cell Lines

In renal medullas during antidiuresis, the extracellular fluid is hyperosmotic because of high concentrations of NaCl and urea. Under those conditions, the cells contain high concentrations of organic osmolytes, namely sorbitol, myo-inositol, glycerophosphorylcholine (GPC), and betaine to balance the extracellular hyperosmolality. These organic osmolytes increase cell osmolality without perturbing the intracellular milieu in ways that would degrade the function of cellular macromolecules. The present study surveyed a number of cell lines for the ability to survive in media with high concentrations of NaCl and/or urea and for the accumulation of organic osmolytes. Of the renal cell lines tested, MDCK, GRB-MAL1, and A6 cells proliferated in hyperosmotic media, but medullary interstitial cells LLC-PK1 and LLC-PK3 did not proliferate, nor did nonrenal HTC-BH cells, MDCK, LLC-PK1, and LLC-PK3 cells contained higher concentrations of myo-inositol, GPC, and betaine when cultured in media containing high NaCl (with or without high urea) and much lower or undetectable levels of these osmolytes when grown in isosmotic media. Sorbitol, and to a lesser extent myo-inositol, were elevated in GRB-MAL1 cells in media hyperosmotic with NaCl but not in isosmotic media. There was less accumulation of organic osmolytes when only urea was added to increase osmolality. Thus the same osmolytes were accumulated by one or another cell line in vitro as were previously found in renal medullas. These cell lines provide models for studying osmolyte accumulation.

scholarly journals Monoclonal antibody defining a molecule possibly identical to the p75 subunit of interleukin 2 receptor.

1989 ◽  
Vol 169 (4) ◽  
pp. 1323-1332 ◽  
Author(s):  
T Takeshita ◽  
Y Goto ◽  
K Tada ◽  
K Nagata ◽  
H Asao ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Interleukin 2 ◽  
High Affinity ◽  
Human T Cell ◽  
Low Concentrations ◽  
High Concentrations ◽  
Dependent Growth ◽  
T Cell Lines

A mouse hybridoma cell line, TU27, producing an mAb was established. TU27 mAb reacted with various human and Gibbon ape T cell lines bearing the IL-2R p75 (IL-2Rp75), but not with cell lines expressing only Tac antigen, IL-2Rp55, and numbers of its binding sites on cell surfaces were similar to those of high-affinity IL-2R. Radioimmunoprecipitation with TU27 mAb defined a molecule with a molecular mass of 75 kD on the surface of IL-2Rp75 bearing cells. TU27 mAb completely blocked IL-2 binding to IL-2Rp75 and to the high-affinity IL-2R but not to IL-2Rp55 composing the low-affinity IL-2R. The IL-2-dependent growth of a human T cell line, ILT-Mat, was significantly inhibited by TU27 mAb only at low concentrations of IL-2, and combination of TU27 mAb and H-31 mAb specific for IL-2Rp55 completely inhibited the cell growth even at high concentrations of IL-2. These data strongly suggest that TU27 mAb is specific for the human IL-2Rp75.

Apoptotic Change Is a Major Reason for Defect in Early Erythroid Development in Cell Line Models for Ribosomal Protein (RP) S19 Deficient Diamond-Blackfan Anemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2840-2840
Author(s):  
Koichi Miyake ◽  
Taiju Utsugisawa ◽  
Johan Flygare ◽  
Thomas Kiefer ◽  
Johan Richter ◽  
...  
Keyword(s):  
Signal Transduction ◽  
B Cells ◽  
Western Blot ◽  
Ribosomal Protein ◽  
Cell Line ◽  
Cell Lines ◽  
Erythroid Differentiation ◽  
Erythroid Progenitor ◽  
Diamond Blackfan Anemia ◽  
Apoptotic Change

Abstract Diamond-Blackfan anemia (DBA) is a congenital red cell aplasia in which 25% of the patients have a mutation in the ribosomal protein (RP) S19 gene. It is not known how the RPS19 deficiency impairs erythopoiesis and proliferation of hematopoietic progenitors. The majority of cases appear to result from an intrinsic disorder of the erythroid progenitor that involves its inability to respond normally to inducers of erythroid proliferation and differentiation. Erythropoietin (Epo) controls the proliferation, differentiation and survival of the erythroid progenitors. However, so far, no receptor-ligand defects have yet been identified. We have established an in vitro models for RPS19 deficient DBA using lentiviral vector mediated doxycycline (Dox) inducible small interfering RNA (siRNA) against RPS19 (Blood102:504a, 2003). To analyze the effect of suppression of RPS19 for erythroid differentiation, we used cytokine dependent TF-1 cell lines (CD34+, CD71+, GFAlow) together with new established cytokine independent K562 cell lines (CD34−, CD71+, GFAhigh). We established two types of cell lines (TF-1A, K562A and TF1B, K562B) using different siRNA against RPS19. Five days after Dox induction, RPS19 protein level was suppressed to 45–55% in TF-1A and K562A, 65–75% in TF1B and K562B compared to control scramble transduced cell lines (TF1S, K562S) by western blot analysis. Suppression of cell growth and colony formation correlated with the suppression level of RPS 19 in TF1A, B and K562A, B cells. In contrast, after stimulation with Epo, glycophorin A expression, measured by flow cytometry, and hemoglobin content (DAF staining) showed suppression of erythroid differentiation only in TF1A and TF1B but not in K562A and K562B. Since Epo induces the stimulation of Jak2 tyrosine kinase which leads to the tyrosine phosphorylation of several proteins including the Epo receptor itself. As a result, different intracellular pathways are activated such as Ras/MAP kinase, phosphatidylinositol 3-kinase and STAT transcription factors. Therefore, we analyzed Epo stimulated signal transduction in these cell lines. However, no abnormal signal transduction could be detected in any of the cell lines. Cell cycle analysis showed that the percentage of cells in the G0/G1 phase increased and apoptotic cells detected by Annexin-V analysis also increased in TF1 (A<B) and K562 (A<B) cells. Western blot analyzed p21 showed that the level of p21 was increased in TF1 (A<B) and K562 (A<B) cells. These results indicate that Epo triggered onset of terminal maturation is intact in RPS19 deficient DBA cell line models. We speculate that apoptotic change by suppressed RPS19 may be one of the major reason for the accelerated loss of erythroid progenitor clonogenicity in RPS19 deficient DBA. These cell lines are useful to determine the mechanisms of RPS19 deficient DBA.

Examining the effects of BET, HDAC2, and EZH2 inhibition on esophageal adenocarcinoma (EAC) cell line proliferation.

2019 ◽  
Vol 37 (4_suppl) ◽  
pp. 68-68
Author(s):  
Kleovoulos Kofonikolas ◽  
Alex M Frankell ◽  
Elizabeth Catherine Smyth ◽  
Rebecca C. Fitzgerald
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Mutant Cell ◽  
Clinical Activity ◽  
Wild Type ◽  
Mutant Type ◽  
Drug Concentrations ◽  
Hematological Cancers ◽  
Before And After ◽  
High Concentrations

68 Background: EACs often carry amplifications in MYC (19%) and mutations in the SWI/SNF complex (13%) which act as oncogenic drivers. Targeting of MYC-amplified tumors via BET inhibition, as well as HDAC2 or EZH2 inhibition in SWI/SNF-mutated tumors, show promise in hematological cancers, and solid tumors, respectively. Herein, we investigate the effects of BET, HDAC2, and EZH2 inhibition on EAC cell line proliferation. Methods: MYC-wild type (n = 5), MYC-amplified (n = 2), SWI/SNF-wild type (n = 6) and SWI/SNF-mutant (n = 1) EAC cell lines that have undergone whole genome sequencing (WGS) were plated at 10% confluency and treated with BET (JQ1), pan-HDAC (SAHA), and EZH2 (EPZ-6438) inhibitors at various concentrations. The CellTiter-Glo cell proliferation assay was used to compare the effects of drug treatment on proliferation between different cell lines and drug concentrations before and after treatment. Results: MYC-amplified EAC cell lines were more sensitive to BET inhibition (Table); GI50 concentrations for JQ1 are comparable to those achieved in melanoma and lymphoma cell lines, cancer types in which clinical activity has been demonstrated. ERBB2 amplified or ERBB2 mutant EAC cells were also sensitive to BET inhibition. HDAC2 inhibition reduced proliferation of both SWI/SNF-wild type and mutant cell lines at high concentrations, whilst all cell lines were resistant to EZH2 inhibition, even at the highest concentrations used. Conclusions: MYC amplified EAC tumors appear to be sensitive to BET inhibition; these results warrant further evaluation in pre-clinical models and clinical trials. HDAC2 and EZH2 inhibition were ineffective in both the SWI/SNF-wild type and mutant type EAC cell lines; these therapies are less likely to provide benefit for EAC patients. [Table: see text]

scholarly journals Influence on Mitochondria and Cytotoxicity of Different Antibiotics Administered in High Concentrations on Primary Human Osteoblasts and Cell Lines

2006 ◽  
Vol 51 (1) ◽  
pp. 54-63 ◽  
Author(s):  
N. Duewelhenke ◽  
O. Krut ◽  
P. Eysel
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Respiratory Chain ◽  
Metabolic Activity ◽  
Lactate Concentration ◽  
Human Osteoblasts ◽  
Extracellular Lactate ◽  
Primary Human Osteoblasts ◽  
High Concentrations ◽  
Local Antibiotic

ABSTRACT Osteomyelitis, osteitis, spondylodiscitis, septic arthritis, and prosthetic joint infections still represent the worst complications of orthopedic surgery and traumatology. Successful treatment requires, besides surgical débridement, long-term systemic and high-concentration local antibiotic therapy, with possible local antibiotic concentrations of 100 μg/ml and more. In this study, we investigated the effect of 20 different antibiotics on primary human osteoblasts (PHO), the osteosarcoma cell line MG63, and the epithelial cell line HeLa. High concentrations of fluoroquinolones, macrolides, clindamycin, chloramphenicol, rifampin, tetracycline, and linezolid during 48 h of incubation inhibited proliferation and metabolic activity, whereas aminoglycosides and inhibitors of bacterial cell wall synthesis did not. Twenty percent inhibitory concentrations for proliferation of PHO were determined as 20 to 40 μg/ml for macrolides, clindamycin, and rifampin, 60 to 80 μg/ml for chloramphenicol, tetracylin, and fluoroquinolones, and 240 μg/ml for linezolid. The proliferation of the cell lines was always less inhibited. We established the measurement of extracellular lactate concentration as an indicator of glycolysis using inhibitors of the respiratory chain (antimycin A, rotenone, and sodium azide) and glycolysis (iodoacetic acid) as reference compounds, whereas inhibition of the respiratory chain increased and inhibition of glycolysis decreased lactate production. The measurement of extracellular lactate concentration revealed that fluoroquinolones, macrolides, clindamycin, rifampin, tetracycline, and especially chloramphenicol and linezolid impaired mitochondrial energetics in high concentrations. This explains partly the observed inhibition of metabolic activity and proliferation in our experiments. Because of differences in the energy metabolism, PHO provided a more sensitive model for orthopedic antibiotic usage than stable cell lines.

Expression and Functional Differences of CBFA2T (ETO, MTG) Gene Family Members in Hematopoiesis.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4208-4208
Author(s):  
Fariborz Mortazavi ◽  
ShriHari Kadkol ◽  
Annette Bruno ◽  
Kristine Baraoidan ◽  
Steven Ackerman ◽  
...  
Keyword(s):  
Gene Family ◽  
Cell Line ◽  
Cell Lines ◽  
Family Members ◽  
Fold Increase ◽  
Erythroid Differentiation ◽  
Macrophage Differentiation ◽  
Human Cd34 ◽  
Cd34 Cells

Abstract The CBFA2T (ETO, MTG) family has three similar family members - CBFA2T1-T3. CBFA2T1 (ETO, MTG8) and CBFA2T3 (ETO2, MTG16) are targeted by chromosomal translocations in acute myeloid leukemia. To better understand the usual hematopoietic function of this gene family, we examined the expression of CBFA2T RNA using RQ-PCR in cell-lines and human CD34+ hematopoietic cells during macrophage and erythroid differentiation. RQ-PCR on extracted RNA was performed with an icyclerQ instrument (Bio-Rad) using the Quantitect SYBR Green RT-PCR kit (Qiagen) and in vitro transcribed RNA to construct standard curves. CBFA2T3 was the most highly expressed family member in human CD34+ cells, the erythro-leukemia line K562, the myeloid line MPD, the T cell line Jurkatt and the B-cell line LCL-11. However, CBFA2T3 expression decreased by >50% during both macrophage and erythroid differentiation of human CD34+ cells. In contrast, CBFA2T1 expression was almost undetectable in human CD34+ cells and all cell lines except K562 but increased more than 20 fold during erythroid (but not macrophage) differentiation of human CD34+ cells. Extrinsic over-expression of CBFA2T1, but not CBFA2T2, significantly increased glycophorin-A and hemoglobin A expression in K562 cells, consistent with a regulatory role for CBFA2T1 in erythroid differentiation. CBFA2T2 (MTGR1) was moderately expressed in human CD34+ cells and all the cell lines and demonstrated a 2.5 fold increase in expression with macrophage differentiation but essentially no change with erythroid differentiation of human CD34+ cells. These findings suggest that despite their similarity, the CBFA2T family members have distinctive regulatory roles in hematopoietic differentiation.

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