DNA Content of Normal and Neoplastic Urothelial Cells

1980 ◽  
Vol 66 (4) ◽  
pp. 445-458 ◽  
Author(s):  
Mathilde E. Boon
Keyword(s):  
Dna Content ◽  
Cell Populations ◽  
Malignant Cells ◽  
Urothelial Cells ◽  
Chromatin Pattern ◽  
Normal Bladder ◽  
Well Differentiated ◽  
Dna Measurements ◽  
Combined Parameters ◽  
Umbrella Cells

In search for suitable parameters to detect neoplastic urothelial cells in Acriflavine-Feulgen-SITS stained specimen we compared the cytofluorometric DNA content with the morphology of normal urothelial cells (bladder scrapings) and neoplastic urothelial cells from grade 1, 2, 3 and 4 tumors. An individual normal urothelial cell could not be distinguished from a grade 1 tumor cell, neither morphologically nor fluorometrically. However, the shape of the histograms of DNA measurements of the cell populations of respectively normal bladder scrapings and grade 1 tumors differs. It is postulated that also morphometry of these cell populations may be of some aid to distinguish well-differentiated neoplastic cells from normal urothelial cells. Seventy-one percent of the morphologically malignant cells in the grade 2, 3 and 4 tumor samples could be identified by applying the combined parameters: high DNA content (> 5 C) and nuclear-cytoplasmic ratio (> 0.5) and all grade 2, 3 and 4 tumor samples contained cells which were objectively classified as malignant. Using the same parameters morphologically malignant cells could be distinguished from normal, polyploid umbrella cells, thus these malignant cells are detectable objectively without using chromatin pattern as parameter.

scholarly journals Measurement of DNA content and of tritiated thymidine and bromodeoxyuridine incorporation by the same cells.

1993 ◽  
Vol 41 (9) ◽  
pp. 1435-1439 ◽  
Author(s):  
P Lin ◽  
D C Allison
Keyword(s):  
Dna Content ◽  
Tritiated Thymidine ◽  
S Phase ◽  
Brdu Incorporation ◽  
Cell Populations ◽  
Feulgen Staining ◽  
Dna Contents ◽  
Incorporated Brdu ◽  
Dna Measurements ◽  
Kinetics Of

We tested a method of measuring DNA content (Feulgen) and tritiated thymidine ([3H]-T) and bromodeoxyuridine (BrdU) incorporation by the same cell. Initial experiments showed that Feulgen hydrolysis denatured the DNA of fixed cells sufficiently to allow detection of incorporated BrdU with monoclonal antibodies. MCa-11 cells were then double-labeled with [3H]-T and BrdU, placed on slides, and Feulgen stained. Next, absorption cytometry was performed to measure the DNA content of randomly selected cells. Feulgen staining and the development and removal of either the [3H]-T or the BrdU grains after DNA measurements did not interfere with subsequent detection of the grains from the other label, and BrdU and [3H]-T can be used reliably in combination for identification of S-phase cells. This method may eventually allow the use of microscope-based image analysis to selectively measure the DNA contents and the BrdU/[3H]-T labeling of non-transformed stromal and cancer cells in solid tumors, thereby providing new insights into the growth kinetics of these heterogeneous cell populations.

Prognostic Implications of Nuclear DNA Content in Head and Neck Cancer

1989 ◽  
Vol 100 (2) ◽  
pp. 95-98 ◽  
Author(s):  
Yang-Chun Guo ◽  
Lawrence Desanto ◽  
Gregory V. Osetinsky
Keyword(s):  
Head And Neck ◽  
Dna Content ◽  
Nuclear Dna ◽  
Clinical Stage ◽  
Nuclear Dna Content ◽  
Survival Advantage ◽  
Prognostic Information ◽  
Diagnosis Of Cancer ◽  
Well Differentiated

The nuclear DNA content was measured in formalin-fixed and deparaffined specimens of 296 oral, pharyngeal, and laryngeal squamous cell carcinomas from patients in whom the clinical outcome was known. One hundred ninety (64%) contained cells with abnormal DNA (DNA aneuploid or tetra/polypoid). Only 32% (60 of 190) of the patients with DNA nondiploid cancers survived 5 years, compared with 49% (52 of 106) of the patients with DNA diploid cancers. When the findings were controlled for clinical stage, patients whose tumors were DNA diploid had a survival advantage at each stage. Histologic grading showed less correlation, because only patients with well-differentiated carcinomas had a survival advantage if their tumors were DNA diploid. These data showed that determination of DNA content in cancers of the head and neck can offer prognostic information not provided by other means and enhance the diagnosis of cancer.

Assessing the differentiation state of cultured bovine urothelial cells: elevated synthesis of stratification-related K5 and K6 keratins and persistent expression of uroplakin I

1990 ◽  
Vol 97 (3) ◽  
pp. 419-432 ◽  
Author(s):  
B. Surya ◽  
J. Yu ◽  
M. Manabe ◽  
T.T. Sun
Keyword(s):  
Feeder Cells ◽  
Serum Free Medium ◽  
Urothelial Cells ◽  
Significant Progress ◽  
Protein Subunit ◽  
A Cell ◽  
Umbrella Cells ◽  
Normal Urothelium ◽  
Made In ◽  
Biochemical Differentiation

Although significant progress has recently been made in culturing mammalian urothelial cells, relatively little is known about their biochemical differentiation. In this paper, we assessed the differentiation state of cultured bovine urothelial cells by analyzing their keratins and a cell surface marker, uroplakin I. Urothelial cells were serially cultured either in a serum-free medium, or in a serum-containing medium in the presence of 3T3 feeder cells, with similar results. Despite their stratified appearance, both normal urothelium and cultured urothelial cells synthesize mainly K8, K18 and K19, keratins that are typically seen in simple epithelia. However, cultured urothelial cells synthesize a greatly increased amount of K5 and K6 keratins, which are usually expressed by stratified epithelia but present only in trace amounts in normal urothelium. These data indicate that, as far as keratin synthesis is concerned, cultured urothelial cells undergo an altered pattern of differentiation towards a more ‘stratified phenotype’; this unusual finding has interesting implications for urothelial evolution. In the meantime, many superficial cells in cultured urothelial colonies make uroplakin I, a 27 × 10(3) Mr protein subunit of the asymmetrical unit membrane (AUM) characteristic of urothelial (superficial) umbrella cells. These results indicate that cultured urothelial cells undergo, at least in part, AUM biogenesis. Cultured urothelial cells thus provide a useful experimental model system for studying certain early steps of AUM formation.

scholarly journals The effect of splenectomy on platelet formation and megakaryocyte DNA content in rats

Blood ◽  
1984 ◽  
Vol 63 (3) ◽  
pp. 593-597 ◽  
Author(s):  
G Tanum ◽  
A Sonstevold ◽  
E Jakobsen
Keyword(s):  
Bone Marrow ◽  
Dna Content ◽  
Direct Action ◽  
Surgical Trauma ◽  
Sham Operation ◽  
Acetylcholinesterase Staining ◽  
Effect Of Splenectomy ◽  
A Minor ◽  
Dna Measurements ◽  
Platelet Formation

Abstract The DNA content of rat bone marrow megakaryocytes (MK) was studied by Feulgen photometry following splenectomy and sham operation, respectively. The DNA measurements were preceded by acetylcholinesterase staining for identification of the 2N-8N MK. The number of 2N-8N MK decreased to minimum values, while the number of 16N- 64N MK increased to maximum values about 4 days following both splenectomy and sham operation. However, the changes were somewhat more pronounced following splenectomy than sham operation. The total MK number did not change significantly. Platelet production, measured by 35S incorporation into platelets, increased during the first 2 days and remained high for 6–7 days, increasing the platelet counts. All values were about normal 30 days after surgery, except for a minor thrombocytosis following splenectomy. The early, highly significant thrombocytosis, following both splenectomy and general surgery, is caused by increased production of platelets due to the surgical trauma. This is caused by a direct action on bone marrow MK by transforming 2N- 8N MK into higher ploidy classes. Lack of splenic platelet pooling may influence the grade and duration of the early thrombocytosis after splenectomy. The late, long-lasting, minor thrombocytosis, which occurs after splenectomy but not after sham operation, can be explained by the removal of the splenic platelet pool.

Determination of nuclear DNA content in fungi using mithramycin: vegetative diploidy in Armillaria mellea confirmed

1983 ◽  
Vol 29 (9) ◽  
pp. 1179-1183 ◽  
Author(s):  
A. L. Franklin ◽  
W. G. Filion ◽  
J. B. Anderson
Keyword(s):  
Dna Binding ◽  
Aspergillus Nidulans ◽  
Dna Content ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Phytopathogenic Fungus ◽  
Armillaria Mellea ◽  
Dna Measurements

Armillaria mellea, a phytopathogenic fungus, is the only member of the Agaricales (Basidiomycetes) whose fertile vegetative phase in nature is thought to be diploid, rather than dikaryotic. To examine the vegetative ploidy of A. mellea, we used the DNA-binding antibiotic, mithramycin, for fluorometry of in situ nuclear DNA. The measurements of nuclear DNA content indicated that strains derived from single basidiospores of A. mellea were haploid and that strains derived from matings of isolates of single spores were diploid. These data confirm the results of earlier genetic experiments, which show haploidy and diploidy in unmated and mated strains, respectively. Nuclear DNA measurements in known haploid and diploid strains of Aspergillus nidulans confirmed the validity of our protocol.

scholarly journals Flow cytometric analysis of small DNA content differences in heterogeneous cell populations: Human amniotic fluid cells

Cytometry ◽  
1984 ◽  
Vol 5 (2) ◽  
pp. 118-123 ◽  
Author(s):  
H. Koch ◽  
T. Bettecken ◽  
M. Kubbies ◽  
D. Salk ◽  
J. W. Smith ◽  
...  
Keyword(s):  
Amniotic Fluid ◽  
Dna Content ◽  
Flow Cytometric Analysis ◽  
Cell Populations ◽  
Human Amniotic Fluid ◽  
Flow Cytometric ◽  
Amniotic Fluid Cells

scholarly journals Beyondcell: targeting cancer therapeutic heterogeneity in single-cell RNA-seq data

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Coral Fustero-Torre ◽  
María José Jiménez-Santos ◽  
Santiago García-Martín ◽  
Carlos Carretero-Puche ◽  
Luis García-Jimeno ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Lines ◽  
Tumour Cell ◽  
Enrichment Score ◽  
Cell Populations ◽  
Drug Selection ◽  
Malignant Cells ◽  
Rna Seq ◽  
Computational Methodology ◽  
Cell Subpopulations

AbstractWe present Beyondcell, a computational methodology for identifying tumour cell subpopulations with distinct drug responses in single-cell RNA-seq data and proposing cancer-specific treatments. Our method calculates an enrichment score in a collection of drug signatures, delineating therapeutic clusters (TCs) within cellular populations. Additionally, Beyondcell determines the therapeutic differences among cell populations and generates a prioritised sensitivity-based ranking in order to guide drug selection. We performed Beyondcell analysis in five single-cell datasets and demonstrated that TCs can be exploited to target malignant cells both in cancer cell lines and tumour patients. Beyondcell is available at: https://gitlab.com/bu_cnio/beyondcell.

scholarly journals The megakaryocyte DNA content and platelet formation after the sublethal whole body irradiation of rats

Blood ◽  
1984 ◽  
Vol 63 (4) ◽  
pp. 917-920 ◽  
Author(s):  
G Tanum
Keyword(s):  
Stem Cell ◽  
Dna Content ◽  
Cell Activation ◽  
Time Lag ◽  
Whole Body ◽  
Cell Compartment ◽  
Applied Dose ◽  
Acetylcholinesterase Staining ◽  
Dna Measurements ◽  
Platelet Formation

The DNA content of rat bone marrow megakaryocytes (MK) was studied by Feulgen photometry, following whole body irradiation with 2 Gy. The DNA measurements were preceded by acetylcholinesterase staining to avoid missing the smaller 2N-8N MK. The number of 2N-8N MK declined immediately following irradiation, whereas the number of 16N-64N MK remained normal for 4 days before decreasing. The number of 2N-8N and 16N-64N MK reached minimum around days 7 and 10, respectively, and thereafter increased to supranormal values at days 14 and 20, respectively. Platelet production, measured by 35S incorporation into platelets, increased during the first 4 days, then decreased to minimum about day 10. A rise to supranormal values was present at day 20. All values were about normal 30 days after exposure. The observed pattern may be explained as follows: Most of the 16N-64N MK survive the applied dose and maintain their ability to produce platelets. Some of the 2N-4N and 8N MK survive irradiation and transform into platelet-producing MK. No influx of cells from the MK stem cell compartment into the MK compartment can be observed before day 7 after irradiation. One explanation for this time lag may be that thrombocytopenia, which does not occur before then, is an essential stimulus for MK stem cell activation.

scholarly journals Epithelial Membrane Antigen, Vimentin, Desmin, Calretinin, E-Cadherin on Cell Block Preparations to Distinguish Well Differentiated Adenocarcinoma from Benign, Reactive, Atypical Mesothelial Cells

2021 ◽  
Vol 10 (18) ◽  
pp. 1302-1308
Author(s):  
Neha Jaiswal ◽  
Jayant Makrande ◽  
Sunita Vagha
Keyword(s):  
Epithelial Membrane Antigen ◽  
Mesothelial Cells ◽  
Definitive Diagnosis ◽  
Cytological Diagnosis ◽  
Cell Block ◽  
Malignant Cells ◽  
Immunohistochemical Markers ◽  
Adenocarcinoma Cells ◽  
Well Differentiated ◽  
E Cadherin

BACKGROUND Inconclusive cytomorphology often results due to failure to distinguish between adenocarcinoma cells from benign, reactive, atypical mesothelial cells in effusion specimens. To resolve such dilemmas, auxiliary techniques like immunohistochemistry were utilised to reach a definitive diagnosis for better treatment and management of patients. We wanted to compare cytodiagnosis achieved on cell block preparations with the cytodiagnosis on conventional smear and perform immunohistochemistry for epithelial membrane antigen (EMA), calretinin, desmin, vimentin and E-cadherin on cell block preparation of the fluids in cases of indistinguishable cytomorphology of adenocarcinoma and reactive, atypical, and benign mesothelial hyperplasia. METHODS The immunohistochemical markers namely EMA, calretinin, vimentin, desmin and Ecadherin were applied on cell blocks employing streptavidin-biotin method. Immunohistochemistry was interpreted by giving scores to the percentage of stained cells. RESULTS EMA and E-cadherin had 100 % sensitivity in diagnosing adenocarcinoma whereas calretinin, vimentin and desmin were 100 % sensitive in diagnosing reactive, atypical mesothelial carcinoma on the cell block preparations. CONCLUSIONS Immunocytochemistry of fluid should be carried out on the cell block preparation where cytological diagnosis on conventional smear and cell block fails to detect malignant cells in the preparation. KEY WORDS Cell Block, Adenocarcinoma, Mesothelial Cells, Immunohistochemistry, EMA, Calretinin, Vimentin, Desmin, E-Cadherin

Clinical significance of SDF-1 isoforms in bladder cancer.

2015 ◽  
Vol 33 (7_suppl) ◽  
pp. 334-334 ◽  
Author(s):  
Marie C Hupe ◽  
Miguel Gosalbez ◽  
Soum D Lokeshwar ◽  
John Shields ◽  
Travis J Yates ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Clinical Significance ◽  
Univariate Analysis ◽  
Differentially Expressed ◽  
Mrna Levels ◽  
Urothelial Cells ◽  
Transcript Levels ◽  
Normal Bladder ◽  
Alternative Mrna Splicing ◽  
Increased Risk

334 Background: Stroma-Derived Factor (SDF)-1 is a ligand for chemokine receptors CXCR4 and CXCR7. The six known SDF-1 isoforms are generated by alternative mRNA splicing. While SDF-1 expression has been detected in various malignancies, only a few studies have reported differential expression of SDF-1 isoforms and its clinical significance. In this study we evaluated the expression three SDF-1 isoforms (α,β,γ) in bladder cancer (BCa). Methods: Using quantitative PCR, mRNA levels of SDF-1α, SDF-1β and SDF-1γ were measured in bladder tissues (normal: 25; BCa: 44) and urine specimens (n=210; normal: 28; benign conditions: 74; BCa: 57, history of BCa (HxBCa): 35, Hx other Ca: 8; other Ca: 8) from consecutive patients. The transcript levels were normalized to β-actin. Correlation between SDF1-isoform transcript levels and clinical outcome (metastasis and disease specific mortality (DSM)) was evaluated in univariate and multivariate analyses. Results: Among SDF-1 isoforms, only SDF-1β mRNA was significantly overexpressed by 2.5-6-fold in BCa tissues when compared to normal bladder tissues. While SDF-1α was expressed in bladder tissues, SDF-1γ expression was undetectable. In univariate analysis, pathologic stage, lymph node positivity for tumor, SDF-1α and SDF-1β transcript levels significantly associated with metastasis and DSM. However, in multivariate analysis, stage (P=0.005) and SDF-1β levels (P=0.017) were independent predictors of metastasis and DSM. In exfoliated urothelial cells, only SDF-1β mRNA levels were differentially expressed and having 91.2% sensitivity and 73.8% specificity for detecting BCa. In patients with HxBCa, elevated SDF-1β levels indicated 4.3-fold increased risk (chi-square: 15.1; P=0.0001; relative risk = 4.3) for developing recurrence within 6-months. Conclusions: SDF-1 isoforms are differentially expressed in bladder tissues and exfoliated urothelial cells. SDF-1β mRNA levels in BCa tissues predict poor prognosis. Further, SDF-1β mRNA levels in exfoliated cells detect BCa with high sensitivity and are potential predictors of future recurrence. Support: Grant NCI/NIH R01 CA-72821-14; Women’s Cancer Association – University of Miami.

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