Flow cytometric analysis of the expression of PCNA during the cell cycle in hela cells and effects of the inhibition of DNA synthesis on it

Cytometry ◽  
1993 ◽  
Vol 14 (8) ◽  
pp. 876-882 ◽  
Author(s):  
Kohsuke Sasaki ◽  
Akira Kurose ◽  
Yoji Ishida
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Hela Cells ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Inhibition Of Dna Synthesis

scholarly journals Coix lacryma-jobi var. ma-yuen Stapf sprout extract induces cell cycle arrest and apoptosis in human cervical carcinoma cells

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Eun Suk Son ◽  
Se-Hee Kim ◽  
Young Ock Kim ◽  
Young Eun Lee ◽  
Sun Young Kyung ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Cell Proliferation ◽  
Cell Cycle Arrest ◽  
Hela Cells ◽  
Flow Cytometric Analysis ◽  
B Cell Lymphoma ◽  
Cell Counting ◽  
Cycle Arrest ◽  
Flow Cytometric

Abstract Background Cervical cancer is the second-leading cause of cancer-related mortality in females. Coix lacryma-jobi L. var. ma-yuen (Rom.Caill.) Stapf ex Hook. f. is the most widely recognized medicinal herb for its remedial effects against inflammation, endocrine system dysfunctions, warts, chapped skin, rheumatism, and neuralgia and is also a nourishing food. Methods To investigate the activity of Coix lacryma-jobi sprout extract (CLSE) on cell proliferation in human cervical cancer HeLa cells, we conducted a Cell Counting Kit-8 (CCK-8) assay. Flow-cytometric analysis and western blot analysis were performed to verify the effect of CLSE on the regulation of the cell cycle and apoptosis in HeLa cells. Results We observed that CLSE significantly inhibited cell proliferation. Furthermore, CLSE dose-dependently promoted cell cycle arrest at the sub-G1/ S phase in HeLa cells, as detected by bromodeoxyuridine (BrdU) staining. The cell-cycle-arrest effects of CLSE in HeLa cells were associated with downregulation of cyclin D1 and cyclin-dependent kinases (CDKs) 2, 4, and 6. Moreover, CLSE induced apoptosis, as determined by flow-cytometric analysis and nuclear DNA fragmentation with Annexin V/propidium iodide (PI) and 4′6′-diamidino-2-phenylindole (DAPI) staining. Induction of apoptosis by CLSE was involved in inhibition of the antiapoptotic protein B-cell lymphoma 2 (Bcl-2) and upregulation of the apoptotic proteins p53, cleaved poly (ADP-ribose) polymerase (PARP), cleaved caspase-3, and cleaved caspase-8. Finally, we observed that CLSE inactivated the phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT) pathways. Conclusions CLSE causes cell cycle arrest and apoptotic cell death through inactivation of the PI3K/AKT pathway in HeLa cells, suggesting it is a viable therapeutic agent for cervical cancer owing to its anticancer effects.

scholarly journals Two-color flow-cytometric analysis of the growth cycle of Plasmodium falciparum in vitro: identification of cell cycle compartments.

1986 ◽  
Vol 34 (12) ◽  
pp. 1651-1658 ◽  
Author(s):  
J D Hare
Keyword(s):  
Cell Cycle ◽  
Plasmodium Falciparum ◽  
Dna Synthesis ◽  
Flow Cytometric Analysis ◽  
Growth Cycle ◽  
Green Fluorescence ◽  
Red Fluorescence ◽  
Color Display ◽  
Flow Cytometric

A previous study (Hare JD, Bahler DW: J Histochem Cytochem 34:215, 1986) has shown that the flow cytometric analysis of acridine-orange-stained Plasmodium falciparum growing in vitro generates a complex two-color display, regions of which correlate with the major morphological stages. In this report, four cell cycle compartments (A-D) are defined by characteristic ratios of red and green fluorescence of cells distributed throughout the erythrocytic cycle as well as by the differential effects of several metabolic inhibitors. The primary characteristic of cells in compartment A is the significant increase in red fluorescence. Inhibition of DNA synthesis by either aphidicolin or hydroxyurea causes the accumulation of cells at the interface between compartments A and B, whereas n-butyrate prevents cells in compartment A from reaching the A-B interface. Cells in compartment A display a small increase in green fluorescence which is independent of DNA synthesis but is enhanced by n-butyrate treatment. Cells in compartment B display a continued increase in red fluorescence coupled with a significant increase in green fluorescence, reflecting the onset of DNA synthesis in compartment B. The transition to compartment C is more abrupt and is associated with a marked increase in green fluorescence and little increase in red fluorescence. Compartment D is characterized by an increase in red fluorescence and a continued rise in green fluorescence. It is postulated that these discontinuities in the two-color display reflect not only changes in the rates of RNA and DNA synthesis but also decondensation of parasite chromatin in compartment A as the organism prepares for DNA synthesis, and re-condensation in compartment D as the newly replicated chromatin prepares for segregation into merozoites. The method described promises to provide a sensitive and rapid technique to study the effects of various factors on the growth cycle of the parasite.

scholarly journals The REC1 Gene of Ustilago maydis, Which Encodes a 3′ → 5′ Exonuclease, Couples DNA Repair and Completion of DNA Synthesis to a Mitotic Checkpoint

Genetics ◽  
1996 ◽  
Vol 143 (1) ◽  
pp. 165-174 ◽  
Author(s):  
Kenan Onel ◽  
Andrew Koff ◽  
Richard L Bennett ◽  
Paul Unrau ◽  
William K Holloman
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Flow Cytometric Analysis ◽  
Ustilago Maydis ◽  
Mitotic Checkpoint ◽  
Temperature Sensitive ◽  
Mrna Levels ◽  
Microtubule Inhibitor ◽  
Flow Cytometric ◽  
Extreme Sensitivity

Abstract Mutation in the REC1 gene of Ustilago maydis results in extreme sensitivity to killing by ultraviolet light. The lethality of the rec1-1 mutant was found to be partially suppressed if irradiated cells were held artificially in G2-phase by addition of a microtubule inhibitor. This mutant was also found to be sensitive to killing when DNA synthesis was inhibited by external means through addition of hydroxyurea or by genetic control in a temperature-sensitive mutant strain defective in DNA synthesis. Flow cytometric analysis of exponentially growing cultures indicated that wild-type cells accumulated in G2 after UV irradiation, while rec1-1 cells appeared to exit from G2 and accumulate in G1/S. Analysis of mRNA levels in synchronized cells indicated that the REC1 gene is periodically expressed with the cell cycle and reaches maximal levels at G1/S. The results are interpreted to mean that a G2-M checkpoint is disabled in the rec1-1 mutant. It is proposed that the REC1 gene product functions in a surveillance system operating during S-phase and G2 to find and repair stretches of DNA with compromised integrity and to communicate with the cell cycle apparatus.

Replication and transcription sites are colocalized in human cells

1994 ◽  
Vol 107 (2) ◽  
pp. 425-434 ◽  
Author(s):  
A.B. Hassan ◽  
R.J. Errington ◽  
N.S. White ◽  
D.A. Jackson ◽  
P.R. Cook
Keyword(s):  
Cell Cycle ◽  
Confocal Microscopy ◽  
Dna Synthesis ◽  
Fluorescent Probes ◽  
Hela Cells ◽  
Nuclear Architecture ◽  
Human Cells ◽  
Dynamic Nature ◽  
Nucleotide Triphosphates ◽  
Transcription Sites

HeLa cells synchronized at different stages of the cell cycle were permeabilized and incubated with analogues of nucleotide triphosphates; then sites of incorporation were immunolabeled with the appropriate fluorescent probes. Confocal microscopy showed that sites of replication and transcription were not diffusely spread throughout nuclei, reflecting the distribution of euchromatin; rather, they were concentrated in ‘foci’ where many polymerases act together. Transcription foci aggregated as cells progressed towards the G1/S boundary; later they dispersed and became more diffuse. Replication was initiated only at transcription sites; later, when heterochromatin was replicated in enlarged foci, these remained sites of transcription. This illustrates the dynamic nature of nuclear architecture and suggests that transcription may be required for the initiation of DNA synthesis.

New thiazacridine agents: Synthesis, physical and chemical characterization, and in vitro anticancer evaluation

2016 ◽  
Vol 36 (10) ◽  
pp. 1059-1070 ◽  
Author(s):  
MBO Chagas ◽  
NCC Cordeiro ◽  
KMR Marques ◽  
MG Rocha Pitta ◽  
MJBM Rêgo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cytotoxic Activity ◽  
Cell Cycle Progression ◽  
Antitumor Agents ◽  
Flow Cytometric Analysis ◽  
Cell Lineage ◽  
Chemical Characterization ◽  
Flow Cytometric ◽  
Physical And Chemical

A series of new thiazacridine agents were synthesized and evaluated as antitumor agents, in terms of not only their cytotoxicity but also their selectivity. The cytotoxicity assay confirmed that all compounds showed cytotoxic activity and selectivity. The new compound, 3-acridin-9-ylmethyl-5-(5-bromo-1 H-indol-3-ylmethylene)-thiazolidine-2,4-dione (LPSF/AA29 – 7a), proved to be the most promising compound as it presents lower half-maximal inhibitory concentration (IC50) values (ranging from 0.25 to 68.03 µM) depending on cell lineage. In HepG2 cells, the lowest IC50 value was exhibited by 3-acridin-9-ylmethyl-5-(4-piperidin-1-yl-benzylidene)-thiazolidine-2,4-dione (LPSF/AA36 – 7b; 46.95 µM). None of the synthesized compounds showed cytotoxic activity against normal cells (IC50 > 100 µM). The mechanism of death induction and cell cycle effects was also evaluated. Flow cytometric analysis revealed that the compounds LPSF/AA29 – 7a and LPSF/AA36 – 7b significantly increased the percentage of apoptotic cells and induced G2/M arrest in the cell cycle progression. Therefore, these new thiazacridine derivatives constitute promising antitumor agents whose cytotoxicity and selectivity properties indicate they have potential to contribute to or serve as a basis for the development of new cancer drugs in the future.

scholarly journals Flow Cytometric Analysis of the Influence of Myosmine on the Cell Cycle

2011 ◽  
Vol 84 (3) ◽  
pp. 355-359 ◽  
Author(s):  
Borislava I. Boteva ◽  
Rada M. Mateva ◽  
Ivan A. Iliev ◽  
Galina A. Gorneva
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric
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