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

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.

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Comparative DNA cytometry of primary and recurrent soft tissue sarcomas.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e22516 ◽

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.

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Prognostic significance of DNA-content in human soft tissue sarcomas

European Journal of Cancer ◽

10.1016/0959-8049(93)91639-3 ◽

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

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

Sarcoma ◽

10.1080/13577149977604 ◽

1999 ◽

Vol 3 (3-4) ◽

pp. 171-175 ◽

Cited By ~ 2

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

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The prognostic value of DNA content measured by image cytometry in soft tissue sarcomas

Annals of Oncology ◽

10.1093/annonc/3.suppl_2.s89 ◽

1992 ◽

Vol 3 ◽

pp. S89-S92 ◽

Cited By ~ 2

Author(s):

H. Pape ◽

Ch. Pöttgen ◽

J.S. Ploem ◽

A.M.J. Van Driel-Kulker ◽

R. Wurm ◽

...

Keyword(s):

Soft Tissue ◽

Dna Content ◽

Prognostic Value ◽

Soft Tissue Sarcomas ◽

Image Cytometry

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

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.e22538 ◽

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.

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Expression of Telomerase Activity and Telomerase RNA in Human Soft Tissue Sarcomas

Archives of Pathology & Laboratory Medicine ◽

10.5858/2000-124-0393-eotaat ◽

2000 ◽

Vol 124 (3) ◽

pp. 393-397

Author(s):

Jinyoung Yoo ◽

Robert A. Robinson

Keyword(s):

Soft Tissue ◽

Soft Tissue Tumor ◽

Tertiary Care ◽

Soft Tissue Sarcomas ◽

Telomerase Activity ◽

Transcriptional Level ◽

Telomerase Rna ◽

Fresh Tissue ◽

Tissue Samples ◽

Tissue Tumor

Abstract Objective.—To investigate whether telomerase is reactivated in soft tissue tumor and whether telomerase activity is regulated at the transcriptional level. Design.—Fresh tissue samples of 24 soft tissue sarcomas were analyzed for telomerase activity by a radioactive polymerase chain reaction–based telomeric repeat amplification protocol assay and for human telomerase RNA (hTR) by an in situ hybridization assay. Setting.—Tertiary care teaching hospital. Patients.—Twenty-four patients with soft tissue tumor were surgically treated. Twelve patients had malignant fibrous histiocytoma, 5 had liposarcoma, 6 had leiomyosarcoma, and 1 had rhabdomyosarcoma. Results.—Telomerase activity was detected in 4 sarcoma samples (17%), all of which were positive for hTR. Expression of hTR was demonstrated in 13 sarcomas (54%), 4 of which were positive for telomerase and 9 of which were negative for telomerase. One (50%) of 2 grade 1 tumors, 9 (50%) of 18 grade 2 tumors, and 3 (75%) of 4 grade 3 tumors showed hTR expression. Conclusions.—The relatively low frequency of telomerase activity in soft tissue sarcomas suggests that telomerase may not play an important role in tumorigenesis in these tumors. Telomerase ladders were demonstrated only in association with tumors expressing hTR. It is noteworthy that half of the patients with grade 1 and 2 tumors expressed hTR, suggesting that telomerase RNA may be useful as a marker for identifying tumor aggressiveness earlier than the conventional histopathologic grading scale.

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