Flow cytometric evaluation of cell-cycle progression in ethyl methanesulfonate and methyl methanesulfonate-exposed P3 Cells: Relationship to the induction of sister-chromatid exchanges and cellular toxicity

1991 ◽  
Vol 18 (2) ◽  
pp. 139-149 ◽  
Author(s):  
S. M. Morris ◽  
O. E. Domon ◽  
L. J. McGarrity ◽  
R. L. Kodell ◽  
D. A. Casciano ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Sister Chromatid ◽  
Cell Cycle Progression ◽  
Sister Chromatid Exchanges ◽  
Methyl Methanesulfonate ◽  
Cycle Progression ◽  
Cellular Toxicity ◽  
Flow Cytometric ◽  
Chromatid Exchanges ◽  
Flow Cytometric Evaluation

scholarly journals Erythrocytes modulate cell cycle progression but not the baseline frequency of sister chromatid exchanges in pig lymphocytes

1997 ◽  
Vol 20 (3) ◽  
pp. 397-403 ◽  
Author(s):  
Miguel A. Reigosa ◽  
Sonia Soloneski ◽  
Carlos F. Garcia ◽  
Marcelo L. Larramendy
Keyword(s):  
Cell Cycle ◽  
Sister Chromatid ◽  
Cell Cycle Progression ◽  
Sister Chromatid Exchanges ◽  
Dependent Manner ◽  
Cycle Progression ◽  
Baseline Frequency ◽  
Chromatid Exchanges ◽  
Human Rbcs

The effect of co-culturing varying concentrations of pig and human red blood cells (RBCs) on the baseline frequency of sister chromatid exchanges (SCEs) and cell-cycle progression in pig plasma (PLCs) and whole blood leukocyte cultures (WBCs) was studied. No variation in SCE frequency was observed between pig control WBC and PLC. Addition of pig and human RBCs to pig PLCs did not modify the baseline frequency of SCEs. On the other hand, cell proliferation was slower in PLCs than in WBCs. The addition of pig or human RBCs to PLCs accelerated the cell-cycle progression of pig lymphocytes. When RBCs were added to PLCs the concentration and time sequence of RBC incorporation affected the cell-cycle progression of swine lymphocytes. When doses of pig or human RBCs equivalent to those present in WBCs were added immediately after PLC stimulation, the cell-cycle kinetics were similar to those of WBCs. Shorter co-incubation periods or a reduction in the dose of RBCs made cell-cycle progression intermediate between PLC and WBC values. Thus, pig and human RBCs modulated the in vitro cell-cycle progression of pig lymphocytes in a time- and dose-dependent manner, and the low baseline frequency of SCEs of pig lymphocytes is independent of the presence or absence of erythrocytes in culture

scholarly journals Anti-Tumor Effect of Rutin on Human Neuroblastoma Cell Lines through Inducing G2/M Cell Cycle Arrest and Promoting Apoptosis

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Hongyan Chen ◽  
Qing Miao ◽  
Miao Geng ◽  
Jing Liu ◽  
Yazhuo Hu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Cell Apoptosis ◽  
Cell Cycle Progression ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Human Neuroblastoma ◽  
Cycle Progression ◽  
Cycle Arrest ◽  
Flow Cytometric

Aims. To further investigate the antineuroblastoma effect of rutin which is a type of flavonoid.Methods. The antiproliferation of rutin in human neuroblastoma cells LAN-5 were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Chemotaxis of LAN-5 cells was assessed using transwell migration chambers and scratch wound migration assay. The cell cycle arrest and apoptosis in a dose-dependent manner was measured by flow cytometric and fluorescent microscopy analyses. The apoptosis-related proteins BAX and BCL2 as well as MYCN mRNA express were determined by RT-PCR analysis. Secreted TNF-αlevel were determined using specific enzyme-linked immunosorbent assay kits.Results. Rutin significantly inhibited the growth of LAN-5 cells and chemotactic ability. Flow cytometric analysis revealed that rutin induced G2/M arrest in the cell cycle progression and induced cell apoptosis. The RT-PCR showed that rutin could decrease BCL2 expression and BCL2/BAX ratio. In the meantime, the MYCN mRNA level and the secretion of TNF-αwere inhibited.Conclusion. These results suggest that rutin produces obvious antineuroblastoma effects via induced G2/M arrest in the cell cycle progression and induced cell apoptosis as well as regulating the expression of gene related to apoptosis and so on. It supports the viability of developing rutin as a novel therapeutic prodrug for neuroblastoma treatment, as well as providing a new path on anticancer effect of Chinese traditional drug.

scholarly journals Diclofenac Induces Apoptosis and Suppresses Diffuse Large B-Cell Lymphoma Proliferation Independent of P53 Status

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5485-5485
Author(s):  
Hesham Hassan ◽  
Michelle Varney ◽  
Bhavana J Dave ◽  
Rakesh K Singh
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Cell Lymphoma ◽  
Flow Cytometric Analysis ◽  
B Cell Lymphoma ◽  
Cycle Progression ◽  
Cycle Arrest ◽  
Flow Cytometric ◽  
Large B Cell Lymphoma ◽  
P53 Status

Abstract Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma (NHL).Despite long-term remission achieved with R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone), relapse occurs in almost one third of the patients. Therefore, there is a need for novel therapeutic targets that are relevant to DLBCL pathogenesis. TP73 gene is a member of the p53 tumor suppressor gene family, which is critical in the regulation of cell cycle and apoptosis. TP73 is located in distal 1p36 chromosomal region that is commonly disrupted in DLBCL. Our previous studies had shown that the differential expression of p73 isoforms correlates with proliferation and apoptosis in DLBCL patient specimens. Furthermore, the experimental modulation of p73 isoforms using expression vectors or siRNA modulates the behavior and regulate the chemotherapeutic response of DLBCL cell line models. Diclofenac is NSAID that has been shown to increase p73 activity, substitute p53 activity and suppress the growth of neuroblastoma. In the present study, we investigated whether diclofenac modulates DLBCL apoptosis and cell cycle progression independent of p53 status. We used cell line models of the GCB-DLBCL (DHL-16 and OCI-Ly7) and the ABC-DLBCL (OCI-Ly3 and Pfeiffer). Because OCI-Ly7 and Pfeiffer have a mutant p53, these cells can model the activity of diclofenac in the presence of mutant p53. We used MTT assay to study the response of the DLBCL cells to various concentrations of diclofenac (25, 50, 100, 150, 200, 250 µM) and at different time points (24, 48, and 72 hours). To decipher the biological effects of diclofenac treatment on DLBCL cells Hema-3 staining was done to visualize morphologic evidence of cell death; propidium iodide-based flow cytometric analysis for cell cycle progression; BrdU incorporation for proliferation; and Annexin-V-Flous flow cytometric analysis for apoptosis. Molecularly, Caspase-GLO assay was used for evaluation of Caspase-3, 7, 8 activity and qRT-PCR was used to estimate the effect of diclofenac treatment on p73 and the p53 family transcriptional target regulating cell cycle (p21) and apoptosis (PUMA, NOXA, BIM, and CD95). Mann-Whitney (for two groups) or ANOVA (for more than two groups) analyses were used to determine the statistical significance for comparisons between different treatment groups. Diclofenac treatment displayed a concentration and duration-dependent suppressive cell proliferative activity against a panel of DLBCL cells independent of p53 status including experimental therapy-resistant models. Diclofenac treatment resulted in cell cycle arrest mainly at the G2/M phase, decreased proliferation, and caused profound cell death (mainly apoptosis and possibly necroptosis). Molecularly, diclofenac treatment was associated with increased activity of caspases- 3, -7 and -8. Increased p53 pathway activity as suggested by induction of expression of a panel of p53 transcriptional targets including the cell cycle regulatory molecule p21 and the pro-apoptotic molecules, PUMA, NOXA, BIM, and CD95, was detected in diclofenac treated DLBCL cells. More importantly diclofenac treatment was associated with enhanced expression of the pro-apoptotic isoforms of the p53 homologue, TAp73. Together, our data demonstrate that clinically non-toxic doses of diclofenac treatment, induces apoptosis and cell cycle arrest of both GCB and ABC-DLBCL cells independent of p53 status and is associated with increased expression of the p73 homologue TAp73. These data highlight the potential of diclofenac as a novel adjuvant therapy in DLBCL. Disclosures No relevant conflicts of interest to declare.

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