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.

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.

Malignant Melanoma in Children and Congenital Melanocytic Nevi: DNA Content and Cell Cycle Analysis by Flow Cytometry

2001 ◽  
Vol 4 (1) ◽  
pp. 73-81 ◽  
Author(s):  
Antonio Alvarez-Mendoza ◽  
Jorge Reyes-Esparza ◽  
Ramon Ruiz-Maldonado ◽  
Eduardo Lopez-Corella ◽  
Norma C. Juarez-Herrera
Keyword(s):  
Risk Factors ◽  
Cell Cycle ◽  
Flow Cytometry ◽  
Malignant Melanoma ◽  
Dna Content ◽  
Melanocytic Nevus ◽  
S Phase ◽  
Congenital Melanocytic Nevus ◽  
M Phase ◽  
Diploid Cells

Malignant melanoma (MM) in children, although a rare neoplasm, can occur within a preexisting congenital melanocytic nevus (CMN). All the potential risk factors for this phenomenon are not well known, but increases in S phase and G2 + M phase of cell cycle, DNA aneuploidy, and cell cycle abnormalities in precursor lesions might be among the risk factors. Using paraffin-embedded tissue, we performed a retrospective analysis of DNA content, aneuploidy, and cell cycle by flow cytometry. Two groups of patients were analyzed: 28 children with CMN who did not developed MM, and 6 patients who further developed MM. In this second group, three patients had four biopsies done before the appearance of MM and in two patients biopsies were done after the appearance of MM. All CMN not associated with MM exhibited diploid cells only, their S phase was 11.5% (± 3.8), and their G2 + M phase was 2.5% (± 2.2). Among those patients who developed MM, 3/6 had an S phase > 15.5 and a G2 + M phase > 2.3 prior to the appearance of MM. Two out of six patients had a tetraploid DNA when MM developed and died with a disseminated MM. They had an S phase > 15.5 and their G2 + M phase was > 2.5. We propose that evaluation of DNA content and cell cycle by flow cytometry is a useful method to supplement biopsy findings in children with CMN who have lesions suspicious of developing a MM.

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.

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.

Apoptosis Was Induced by Casticin in Acute Myelocytic Leukemia Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3991-3991
Author(s):  
Jie Jin ◽  
Jia-Kun Shen ◽  
Hua-ping Du ◽  
Min Yang ◽  
Yun-Gui Wang
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cell Death ◽  
Leukemic Cell ◽  
K562 Cells ◽  
Acute Myelocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Cycle Arrest ◽  
M Phase ◽  
Ic50 Values

Abstract Casticin, a component from Vitex rotundifolia wich was widely used as an anti-inflammatory agent in Chinese traditional medicine, was reported to have anti-tumor activities in lung cancer and breast cancer. There are yet no reports on roles against acute myelocytic leukemia (AML). This study aims to elucidate the anti-leukemic activity of casticin on AML cells. We investigated the efficient efficacy and the mechanisms by which casticin triggers cell death in AML cells by analyzing cell cycle perturbations, apoptosis-related marker expression. Cell viability was measured by MTT method; apoptosis and cell cycle arrest were determined by flow cytometry and AV-PI assay. Western blot was performed to measure the apoptosis-related marker. Concentration-dependant cell deaths were observed in AML cell lines including K562, U937 and THP-1, with IC50 values of 24h (hours) being 47.4μM, 67.8μM and 61.7μM, respectively. Time-dependant cell deaths were also observed. At the concentration of 20μM casticin, 45.7%, 76.1% and 80.9% of K562 cells were inhibited at 24h, 48h and 72h, respectively; 24.7%, 30% and 61% of U937 cells were inhibited at 24h, 48h and 72h, respectively; while for THP-1, 29%, 41.8% and 53.9% were inhibited at 24h, 48h and 72h, respectively. Apoptosis was found using AV-PI staining by flow cytometry analysis. We observed an obvious G2/M phase increase prolongation in casticin treated K562 cells. BThe distribitions of G2/M phase were 2.9%, 33.6%, 75.3%, 54.9%, 29.7% and 27.0% in K562 cells after treated by 20μM casticin for 0h, 6h, 12h, 24h, 36h and 48h, respectively. Furthermore, apoptosis-related proteins, PARP and caspase 3, were cleaved in casticin treated K562 cells. Taken together, these results demonstrated that casticin can induce leukemic cell death through apoptosis, suggesting that casticin could be a promising therapeutic agent against acute myeloid leukemia.

Aptamer TY04 Inhibits Multiple Myeloma Cell Growth Via Cell Cycle Arrest

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5390-5390
Author(s):  
Jing Liu ◽  
Hong-Juan Dai ◽  
Bian-Ying Ma ◽  
Jian-Hui Song ◽  
Hui-yong Chen ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Cell Growth ◽  
Cell Lines ◽  
Myeloma Cell ◽  
Multiple Myeloma Cell ◽  
Cycle Arrest ◽  
M Phase ◽  
Target Molecules

Abstract Multiple myeloma (MM), also known as plasma cell myeloma, is characterized by accumulation of clonal plasma cells in the bone marrow and overproduction of monoclonal immunoglobulin (Ig) in the blood or urine. MM accounts for approximately 10% of all hematologic malignancies. Despite recent advances in the understanding and treatment of this disease, MM remains an incurable disease in the vast majority. With conventional chemotherapy, the 5-year median survival rate for MM patients is approximately 25%. Aptamers are single-stranded RNA or DNA sequences that bind to target molecules with high affinity and specificity. Compared with antibodies, aptamers have unique advantages including easy chemical synthesis and modification, low toxicity, lack of immunogenicity, and rapid tissue penetration, Based on these advantages, aptamers show great potential for therapeutic application. The aptamer TY04 is a single-stranded DNA (ssDNA) generated by a method named cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX), We found TY04 strongly inhibited the growth of multiple myeloma cell lines including MM1.S, NCI-H929, KM3 and OPM2,The concentration of TY04 to inhibit 50% cell growth (IC50) on MM1.S was 3.89 μM. In contrast, TY04 had no effect on the growth of non-tumor cell lines — immortal B lymphoblastoid cell lines. Next, we used MM1.S cell line as the model to study the mechanism of TY04 anti- multiple myeloma. Flow cytometry analysis showed that TY04 with the sequence specifically bind to MM1.S cells when compared with unselected ssDNA library control. To investigate whether the target molecules of TY04 are membrane proteins on cell surface, MM1.S cells were treated with trypsin and proteinase k for 2 or 10 minutes before incubation with TY04. The result revealed that TY04 lost partly recognition ability on treated cells, indicating that the target molecules were most likely membrane proteins. Furthermore, we evaluated the cell cycle distribution of MM1.S after TY04 treatment. We found that TY04 significantly caused cell-cycle arrest in G2/M phase. The percentage of G2/M phase cells increased from 30.1±1.56 to 53.2±6.36. To identify the underlying molecular mechanism, G2/M-related proteins were assayed by flow cytometry. Following TY04 treatment, a concomitant inhibition of ERK1/2, cyclin B, CDK1 and γ-tubulin expression occurred. Meanwhile, human cell cycle PCR array was used to analyze the expression of 84 genes key to cell cycle regulation in TY04-treated MM1.S cells. Our results indicated that aptamer TY04 decreased the genes expression of CCNB1, CCNB2, BIRC5, BRCA1 and CCNH, which were involved in the progress of G2/M phase. All these results are significant in that they provide a framework for further exploring the use of TY04 as a novel anti-multiple myeloma agent. Disclosures: No relevant conflicts of interest to declare.

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 Efek Sinergis Kombinasi Ekstrak Air Akar Batu (Gerrardanthus Macrorhizus) dengan Doxorubicin pada Sel Kanker Payudara T47D

2018 ◽  
Vol 46 (3) ◽  
pp. 183-190
Author(s):  
Sari Haryanti
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Cytotoxic Effect ◽  
Clinical Results ◽  
Morphological Alteration ◽  
T47d Cells ◽  
Natural Agents ◽  
Chemotherapy Agent ◽  
M Phase ◽  
Apoptotic Induction

Doxorubicin is one of the most widely used chemotherapy agents in cancer therapy. The clinical results of doxorubicin are limited by cardiotoxic side effects which correlates with dose. Combination of doxorubicin with natural agents is a promising strategy for reducing its therapeutic doses as well as decreasing cardiotoxic effects. This study was conducted to investigate cytotoxic activity of caudex Gerrardanthus macrorhizus aqueous extract and its combination with doxorubicin on T47D cells. Dried caudex powder was extracted by infusion method, evaporated in oven 400C to get dried extract (GM). The MTT assay was performed to determine cytotoxic effect, either alone or in combination. Flow cytometry is used to observe cell cycle profile and apoptotic induction. GM alone did not exhibit cytotoxic effects but caused morphological alteration in T47D cells. Nevertheless, GM 40 μg/mL was able to improve cytotoxic effect of doxorubicin to 51%. Combination of doxorubicin 3 nM and 40 μg/mL GMA inhibited cell cycle in G2/M phase. This combination also resulted apoptotic induction, compared to untreated cell and each single treatment. Based on the results, caudex G. macrorhizus is potential to be further investigated as a co-chemotherapy agent with doxorubicin. Abstrak Doxorubicin adalah salah satu agen kemoterapi yang digunakan secara luas dalam terapi kanker. Hasil klinis doxorubicin dibatasi oleh kardiotoksisitas yang berkorelasi dengan besaran dosis. Kombinasi doxorubicin dengan bahan alam merupakan strategi yang menjanjikan untuk mengurangi dosis terapetik doxorubicin sehingga dapat menurunkan efek samping kardiotoksik. Penelitian ini dilakukan untuk mengkaji aktivitas sitotoksik ekstrak air  caudex Gerrardanthus macrorhizus dan kombinasinya dengan doxorubicin pada sel T47D. Serbuk caudex G. macrorhizus (GM) kering diekstraksi dengan metode infusa, kemudian diuapkan dalam oven 400C. Uji MTT dilakukan untuk menguji efek sitotoksik ekstrak, baik tunggal maupun kombinasi dengan doxorubicin. Flow cytometry digunakan untuk mengetahui profil siklus sel dan induksi apoptosis. Hasil penelitian menunjukkan ekstrak tunggal tidak memberikan efek sitotoksik namun mengakibatkan perubahan morfologi pada sel T47D. Ekstrak GM 40 μg/mL mampu meningkatkan efek sitotoksik doxorubicin 3 nM hingga 51%. Kombinasi doxorubicin 3 nM dan ekstrak 40 μg/mL menghambat siklus sel pada fase G2/M dan meningkatkan induksi apoptosis, dibandingkan kontrol sel dan masing-masing perlakuan tunggalnya. Berdasarkan hasil penelitian ini, caudex G. macrorhizus berpotensi untuk diteliti lebih lanjut sebagai agen ko-kemoterapi dengan doxorubicin.  

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