Comparative DNA cytometry of primary and recurrent soft tissue sarcomas.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e22516-e22516
Author(s):  
Irina Dashkova ◽  
Larisa N. Vashchenko ◽  
Oleg Ivanovich Kit ◽  
Inna A. Novikova ◽  
Ekaterina Komarova ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Soft Tissue ◽  
Dna Content ◽  
Soft Tissue Sarcomas ◽  
Literary Analysis ◽  
Proliferation Index ◽  
Cell Distribution ◽  
Dna Cytometry ◽  
Flow Cytofluorometry ◽  
M Phase

e22516 Background: Soft tissue sarcomas (STS) are aggressive tumors with a high degree of recurrence. Radical resection within healthy tissues allows to reduce the recurrence percentage to 25-30% without subsequent therapy. The literary analysis has shown that the study of various biological properties of primary and recurrentsoft tissue tumorsis being conducted. However, currently there is a lack of information to understand the reasons for STS recurrence. The goal of investigation was to reveal the distinctive features of the DNA content and cell distribution in the phases of the cell cycle of recurrent STS. Methods: DNA cytometry in the tumor tissue of 30 primary soft tissue sarcomas and 30 STS recurrences was carried out using the method of flow cytofluorometry. The tumor ploidy and cell distribution in the cell cycle phases were analyzed. Results: A number of differences in the DNA cytometric parameters of primary and recurrent STS have been revealed, they include: an increase in the proportion of aneuploid tumors in case of recurrence, the number of tumors with DNA index within the mitotic cycle, an increase in the proportion of cells in G2+M- phase of diploid and aneuploid tumors and a decrease in S- phase of aneuploid ones. It has been shown that with a G2 differentiation degree, the proportion of cells in G2+M, S- and proliferation index of recurrent STS is significantly lower than the primary parameters. An increase in the proportion of cells in G2+M- phase and a decrease in the rate of proliferation of recurrent STS, depending on the stage, are shown only in case of stage III. Conclusions: The revealed features of DNA content and cell cycle of tumor cells of soft tissue sarcomas will allow to approach to understanding of biological bases of recurrence of this malignant disease.

Comparative analysis of cell cycle parameters in primary and recurrent soft tissue sarcomas.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e22538-e22538
Author(s):  
Inna Arnoldovna Novikova ◽  
Evgeniya M. Nepomnyashchaya ◽  
Timur Aliev ◽  
Elena Yurievna Zlatnik ◽  
Olesya N. Selyutina ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Comparative Analysis ◽  
Soft Tissue ◽  
Dna Content ◽  
Soft Tissue Sarcomas ◽  
Disease Stage ◽  
Primary Tumors ◽  
Recurrent Tumors ◽  
M Phase

e22538 Background: The purpose of the study was to determine DNA content and distribution of cells in cell cycle phases by flow cytometry in patients with primary and recurrent soft tissue sarcomas. Methods: 60 patients with soft tissue sarcomas (STS) were recruited: 30 with primary tumors and 30 with recurrent ones. Mean age of patients with primary STS was 56±5.4 years, with recurrent STS – 55±6.7 years. DNA content was determined using the BD Facs Cantoo II flow cytometer with CycleTEST PLUS DNA Reagent Kit (Becton Dickinson). The data were processed using ModFit LT program. Results: Comparative analysis of the cell cycle kinetics showed an increase in the percentage of cells in G2+M phase by 2 times in diploid and by 2.1 times in aneuploid recurrent tumors in comparison with primary ones (1.8±0.5% vs. 0.9±0.1% for diploid tumors; 5.4±2.2% vs. 2.6±0.7% for aneuploid tumors). An increase in the percentage of aneuploid tumors was found in recurrent G2 and G3 tumors (from 50% in primary to 66.7% in recurrent G2 tumors and from 63.25% in primary to 85% in recurrent G3 tumors). Mean content of aneuploid cells in recurrent G2 tumors was 2.2 times higher (p≤0.05), while the differences in primary and recurrent G3 tumors were not significant. The percentage of aneuploid tumors depended on the disease stage and increased in stages IIb and III in recurrent tumors, compared to primary ones (from 37.5% to 71.4% in recurrent st. IIb; and from 65% in primary st. III to 72.7% in recurrences) (p≤0.05). Conclusions: DNA analysis by flow cytometry demonstrated a high biologic potential of both primary and recurrent tumors. Some values in the mitotic cycle in recurrent tumors were probably associated with adjuvant therapy, as well as influenced by the coefficient of two parameters – the percentage of cells in G2+M phase and the cell loss factor determining a high malignant potential of these tumors.

scholarly journals DNA-cytometric characteristics of recurrent soft tissue sarcomas

2018 ◽  
Vol 99 (3) ◽  
pp. 415-420
Author(s):  
E M Nepomnyashchaya ◽  
E P Ul'yanova ◽  
I A Novikova ◽  
O N Selyutina ◽  
E Yu Zlatnik ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Soft Tissue ◽  
Soft Tissue Sarcomas ◽  
Tumor Grade ◽  
Cell Loss ◽  
Biological Potential ◽  
M Phase ◽  
Poorly Differentiated ◽  
G0 Phase ◽  
Well Differentiated

Aim. To determine the content of deoxyribonucleic acid (DNA) and distribution of cells in mitotic phases in patients with recurrent soft tissue sarcomas for the assessment of malignancy of the process. Methods. Tumor tissues of patients with recurrent soft tissue sarcomas were studied. Research methods included histological, DNA-cytometric and statistical methods. Results. Proliferative activity and proliferative index of recurrent sarcomas differed depending on the tumor grade and stage. Differences in the number of diploid, aneuploid and polyploid cells were determined in each group and between the groups depending on the cell cycle phases. Cell cycle parameters were as following: 100% of G1 (well-differentiated) cancer were diploid, as well as 33.3% of G2 (moderately differentiated) and 15% of G3 (poorly differentiated) tumors. Aneuploid tumors prevailed in G2 and G3, the ratio of which was 66.7 and 85%, respectively. The analysis of kinetic parameters of the cell cycle allowed establishing a decrease in the number of cells in G1/G0 phase of the cell cycle from G1 to G3, which was accompanied by a statistically significant increase in the proportion of cells in S-phase (p ˂0.05). Conclusion. The DNA-cytometric study of cell cycle parameters showed high biological potential of recurrent soft tissue sarcomas, which was determined by two indices - the proportion of cells in G2+M-phase and the cell loss factor; 100% of well-differentiated (G1) tumors, 33.3% of moderately differentiated (G2) and 15% of poorly differentiated (G3) tumors were diploid; aneuploid tumors prevailed in G2 and G3.

Prognostic Implications of p53 Nuclear Overexpression and High Proliferation Index of Ki-67 in Adult Soft-Tissue Sarcomas

1994 ◽  
Vol 86 (7) ◽  
pp. 549-554 ◽  
Author(s):  
Marija Drobnjak ◽  
Esther Latres ◽  
Daphna Pollack ◽  
Martin Karpeh ◽  
Maria Dudas ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Soft Tissue Sarcomas ◽  
Proliferation Index ◽  
Ki 67 ◽  
Prognostic Implications

Cellular DNA content and prognosis of high-grade soft tissue sarcoma: the Scandinavian Sarcoma Group experience.

1990 ◽  
Vol 8 (3) ◽  
pp. 538-547 ◽  
Author(s):  
T A Alvegard ◽  
N O Berg ◽  
B Baldetorp ◽  
M Fernö ◽  
D Killander ◽  
...  
Keyword(s):  
Risk Factors ◽  
Risk Factor ◽  
Soft Tissue ◽  
Dna Content ◽  
Nuclear Dna ◽  
Single Agent ◽  
Soft Tissue Sarcomas ◽  
Nuclear Dna Content ◽  
High Grade ◽  
Dna Aneuploidy

The nuclear DNA content of 148 high-grade soft tissue sarcomas of the extremities and trunk was determined by flow cytometry, using tumor material from paraffin-embedded tissue. The patients were part of a prospective randomized clinical trial on the efficacy of adjuvant single-agent chemotherapy with doxorubicin. Chemotherapy did not improve the metastasis-free survival (MFS). After a median follow-up time of 48 months (range, 2 to 97), a multivariate analysis of prognostic factors for developing metastatic disease was performed. DNA aneuploidy was found to be an independent prognostic risk factor in addition to histologic malignancy grade IV, intratumoral vascular invasion, tumor size over 10 cm, and male sex. Patients with none or one risk factor had a 5-year MFS of 79%, with two risk factors 65%, with three risk factors 43%, and with four and five risk factors 0%. About one half (78 of 148) of the patients with three factors or less belonged to a group with a MFS over 60%. The combination of different risk factors, including DNA aneuploidy, seems to be a useful prognostic model for soft tissue sarcomas, which could be of value to select high-risk patients for further trials with adjunctive therapy.

Prognostic significance of DNA-content in human soft tissue sarcomas

1993 ◽  
Vol 29 ◽  
pp. S183
Author(s):  
W. Budach ◽  
B. Socha ◽  
V. Budach ◽  
C. Streffer ◽  
H Sack
Keyword(s):  
Soft Tissue ◽  
Dna Content ◽  
Prognostic Significance ◽  
Soft Tissue Sarcomas ◽  
Human Soft Tissue

scholarly journals Recurrent DMD Deletions Highlight Specific Role of Dp71 Isoform in Soft-Tissue Sarcomas

Cancers ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 922 ◽  
Author(s):  
Olivier Mauduit ◽  
Vanessa Delcroix ◽  
Tom Lesluyes ◽  
Gaëlle Pérot ◽  
Pauline Lagarde ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Cell Lines ◽  
Cell Cycle Progression ◽  
Soft Tissue Sarcomas ◽  
Metastatic Progression ◽  
Gene Encoding ◽  
M Phase ◽  
Synovial Sarcomas ◽  
Dmd Gene

Soft-tissue sarcomas (STS) are rare tumors whose oncogenesis remains unknown and for which no common therapeutic target has yet been identified. Analysis of 318 STS by CGH array evidenced a frequent deletion affecting the DMD gene (encoding dystrophin isoforms) in 16.5% of STS, including sarcomas with complex genomics, gastrointestinal tumors (GIST), and synovial sarcomas (SS). These deletions are significantly associated with metastatic progression, thus suggesting the role of DMD downregulation in the acquisition of aggressive phenotypes. We observed that targeted deletions of DMD were restricted to the 5’ region of the gene, which is responsible for the transcription of Dp427. Analysis of STS tumors and cell lines by RNA sequencing revealed that only the Dp71 isoform was widely expressed. Dp427 depletion had no effect on cell growth or migration. However, Dp71 inhibition by shRNA dramatically reduced the cell proliferation and clonogenicity of three STS cell lines, likely by altering the cell cycle progression through the G2/M-phase. Our work demonstrates that DMD deletions are not restricted to myogenic tumors and could be used as a biomarker for metastatic evolution in STS. Dp71 seems to play an essential role in tumor growth, thus providing a potential target for future STS treatments.

Altered Expression of G1 Regulatory Proteins in Human Soft Tissue Sarcomas

2002 ◽  
Vol 126 (5) ◽  
pp. 567-573 ◽  
Author(s):  
Jinyoung Yoo ◽  
Sonya Y. Park ◽  
Seok Jin Kang ◽  
Sang In Shim ◽  
Byung Kee Kim
Keyword(s):  
Cell Cycle ◽  
Soft Tissue ◽  
Cyclin D1 ◽  
Soft Tissue Sarcomas ◽  
Tumor Grade ◽  
Regulatory Proteins ◽  
Nuclear Accumulation ◽  
Cyclin D ◽  
P53 Pathway ◽  
Altered Expression

Abstract Context.—Soft tissue sarcomas constitute a heterogeneous group of tumors for which tumorigenesis is not fully understood. Altered cell-cycle regulation may underlie the development and/or progression of human malignancies. However, data concerning the occurrence of cell-cycle aberrations in soft tissue sarcomas are very limited. Objectives.—To detect the abnormal features of cell-cycle regulatory proteins in soft tissue sarcomas and to determine the potential role of these proteins in clinical behavior. Design.—The p53 and Rb–cyclin D pathways were investigated by immunohistochemical studies of p53, mdm2, pRb, p16, cyclin D1, and cdk4 proteins, respectively. Results.—Of the 67 sarcomas analyzed, nuclear accumulation of p53 was detected in 25 samples (37%), and overexpression of mdm2 was found in 16 samples (24%). Both p53 and mdm2 expression correlated with tumor grade. Abnormalities involving the Rb–cyclin D pathway were identified in all of the tumors by the altered expression of either pRb (72%) or p16 (94%). Fourteen (21%) and 64 (96%) cases demonstrated cyclin D1 or cdk4 expression, respectively. Overexpression of cyclin D1 showed an association with pRb and p53. There was no correlation between pRb, p16, cyclin D1, or cdk4 and tumor grade or relapse. Conclusion.—Disturbance in the cell-cycle regulatory system involving the p53 pathway and the Rb–cyclin D pathway is relatively frequent in soft tissue sarcomas and may be a contributing factor in the tumorigenesis of these tumors. The alterations in the Rb–cyclin D pathway probably constitute an early event, whereas the abnormalities in the p53 pathway seem to be involved in tumor progression. It is noteworthy that cyclin D1 may play a key role in linking both pathways.

DNA content parameters of paraffin-embedded soft tissue sarcomas: Optimization of retrieval technique and comparison to fresh tissue

Cytometry ◽  
1993 ◽  
Vol 14 (3) ◽  
pp. 327-333 ◽  
Author(s):  
Mark M. Zalupski ◽  
Zosia Maciorowski ◽  
James R. Ryan ◽  
John F. Ensley ◽  
Mohamed E. Hussein ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Dna Content ◽  
Soft Tissue Sarcomas ◽  
Fresh Tissue ◽  
Retrieval Technique

scholarly journals The Clinical Value of Flow Cytometric DNA Content Analysis in Patients with Soft Tissue Sarcomas

Sarcoma ◽  
1999 ◽  
Vol 3 (3-4) ◽  
pp. 171-175 ◽  
Author(s):  
Mustafa Samur ◽  
Ali Pamir ◽  
Hakan Akbulut ◽  
Selim Erekul ◽  
Yener Sağlik ◽  
...  
Keyword(s):  
Content Analysis ◽  
Soft Tissue ◽  
Dna Content ◽  
Soft Tissue Sarcomas ◽  
Clinical Value ◽  
Flow Cytometric

scholarly journals Multiparametric Cell-Based Assay for the Evaluation of Transcription Inhibition by High-Content Imaging

2013 ◽  
Vol 18 (5) ◽  
pp. 556-566 ◽  
Author(s):  
Raquel Torres-Guzmán ◽  
Shaoyou Chu ◽  
Juan A. Velasco ◽  
María José Lallena
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Dna Content ◽  
Human Cancer ◽  
Linear Regression Analysis ◽  
Proliferation Index ◽  
Transcription Inhibition ◽  
High Content Imaging

Loss of normal cell cycle regulation is a hallmark of human cancer. Cyclin-dependent kinases (CDKs) are key regulators of the cell cycle and have been actively pursued as promising therapeutic targets. Likewise, members of the CDK family are functionally related to transcriptional modulation, a molecular pathway suitable for therapeutic intervention. We used a set of 2500 compounds in the U2OS cell line to evaluate its effect in the cell division process. Interestingly, out of this analysis, we identified a subpopulation of compounds that are able to inhibit RNA polymerase activity, thus interfering with gene transcription processes. After this finding, we developed, validated, and fully automated a multiparameter high-content imaging (HCI) assay to measure phosphorylation of the RNA polymerase II carboxyl terminal domain (pCTD). Simultaneously, we measured both the DNA content and cell proliferation index in the treated cells. The linear regression analysis comparing the IC50 for pCTD and the 4N EC50 for DNA content or IC50 for cell proliferation showed an excellent agreement ( r2 = 0.84 and r2 = 0.94, respectively). Our results confirm that this method allows discriminating between cell cycle and transcription inhibition and confirms HCI as a powerful technology for the identification of compounds with an effective and selective pathway phenotype.

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