CD4+ Lymphocytes in Asymptomatic HTLV-1 Carriers Present Cell Cycle Arrest in G0/G1-Phase

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5377-5377 ◽  
Author(s):  
Mari Cleia Martins Rodrigues Ferreira ◽  
Renata Kikuchi Foltran ◽  
Rodrigo Santucci ◽  
Luis Alberto de Padua Covas Lage ◽  
Debora Levy ◽  
...  
Keyword(s):  
Cell Cycle ◽  
S Phase ◽  
High Rate ◽  
G1 Phase ◽  
Control Group ◽  
Dna Index ◽  
Asymptomatic Carriers ◽  
M Phase ◽  
And Control ◽  
Cd4 Lymphocytes

Abstract Introduction: Adult T-cell leukemia/lymphoma (ATLL) is an aggressive and incurable disease caused by human T-lymphotropic virus 1 (HTLV-1) that infects CD4+ and CD8+ lymphocytes, but most commonly the malignant cell present a CD4+ phenotype. However, clonal expansion and cell cycle abnormalities have been demonstrated in CD4+ and in CD8+ lymphocytes of HTLV-1 carriers. Objectives: This study compared DNA content and G0/G1, G2/M and “S”-phases of CD4+ and CD8+ lymphocytes among asymptomatic HTLV-1 carriers, ATLL and health subjects. Methods: Werestudied 38 HTLV-1 carriers, 20 ATLL and 35 health subjects pared by sex and age at the Hematology Department of the Faculty of Medicine, University of São Paulo. Peripheral blood mononuclear cells (PBMCs) were isolated on Ficoll-Paque® and lymphocytes subtypes were obtained by positive selection in a magnetic column. Cell-cycle distribution and DNA index (DI) was assessed by flow cytometry after propidium iodide staining. Results: In ATLL, themedian age was 53.5 years (24 to 72) and 50% were female, in HTLV-1 carriers was 55.5 years (33 to 80) with 63.2% of female and in control group was 50 years (24 to 80) with 54.3% of female. In the CD4+ lymphocyte a % of cells in G0/G1 (98.32%) in HTLV-1 carriers was higher than in control group (97.14%) (p=0.041) and in ATLL (97.25%) (p=0.023). S-phase was not statistically different in asymptomatic carriers (0.34%) and control group (0.63%) (p=0.073), but was higher in ATLL (1.80%) than in asymptomatic carriers (0.34%) (p<0.001) and in control group (0.63%) (p=0.02). G2/M-phase was not significantly different among all groups (p=0.960) (Table 1). The CD4+ lymphocytes were aneuploidy in 39.5% (18.4% DI > 1.05 and 21.5% < 0.95) of asymptomatic carriers and in 26.7% (20% > 1.05 and 6.7% < 0.95) of ATLL patients (p=0.557). All control groups were diploid. Table 1.Comparison of the cell cycle by flow cytometry of T lymphocytes CD4+CD4+ cellsAsymptomatic carriersATLLControl groupp-ValueG0/G1mean(dp)97.78 (2.182)95.69 (3.557)96.55 (2.964)0.0351º; median;3ºq97.03;98.32;99.6491.40;97.25;98.3295.01;97.14;98.64G2/Mmean(dp)1.55(1.848)1.91(2.798)2.03(2.902)0.961º; median;3ºq0.00;0.88;2.670.12;0.99;1.990.00;0.56;2.97S-phasemean(dp)0.68(1.207)2.80(3.372)1.43(1.780)0.0031º; median;3ºq0.00;0.34;0.650.65;1.80;3.510.04;0.63;2.55 In CD8+ there was no found significantly difference in whole groups for G0/G1-phase (p=0.138) and G2/M-phase (p=0.374). ATLL presented higher S-phase (median 1.54%) than asymptomatic carriers (median 0.45%) (p=0.003) and control group. S-phase in asymptomatic carriers was not significantly different in comparison to control group (p=0.712). CD8+ were aneuploidy in 23.7% (5.3% DI > 1.05 and 18.4% < 0.95) of asymptomatic carriers and in 21% (10.5% > 1.05 and 10.5% < 0.95) of ATLL (p=0.603). In ATLL the median of DI was 1.01 (1.0; 1.05) in CD4+ and higher than in CD8+ median 0.99 (0.98; 1.0) (p=0.007). Aneuploidia was seen in 47.7% of ATLL, 26,7% (20% DI > 1.05 and 6,7% < 0.95) in CD4+ and 21,0% in CD8+ (10,5% > 1.05 and10,5% < 0.95) (p=0.625). Figure 1: Dna index of CD4+ and CD8+. Aneuploidia was found in HTLV I carriers in both CD4+ and CD8+. Figure 1:. Dna index of CD4+ and CD8+. Aneuploidia was found in HTLV I carriers in both CD4+ and CD8+. Figure 2. Comparison of DI between CD4+ and CD8+ of asymptomatic carriers and ATLL Figure 2. Comparison of DI between CD4+ and CD8+ of asymptomatic carriers and ATLL Figure 3 Figure 3. Conclusion: We demonstrated for the first time “in vivo” that asymptomatic HTLV-1 carriers display cell cycle arrest in G0/G1-phase in CD4+ lymphocytes and high rate of aneuploidia in both CD4+ and CD8+. ATLL showed high rate of hiperdiploidia in CD4+ and hipodiploidia in CD8+ and high rate of S-phase in CD4+. Genetic instability and proliferative disturbs are a hallmark not only in ATLL but also in HTLV-1 carriers and in both CD4+ and CD8+ lymphocytes. Disclosures No relevant conflicts of interest to declare.

scholarly journals Fluctuation in radioresponse of HeLa cells during the cell cycle evaluated based on micronucleus frequency

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hiroaki Shimono ◽  
Atsushi Kaida ◽  
Hisao Homma ◽  
Hitomi Nojima ◽  
Yusuke Onozato ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Hela Cells ◽  
Time Lapse ◽  
S Phase ◽  
G1 Phase ◽  
M Phase ◽  
G2 Phase ◽  
The Difference ◽  
Time Lapse Imaging ◽  
First Time

AbstractIn this study, we examined the fluctuation in radioresponse of HeLa cells during the cell cycle. For this purpose, we used HeLa cells expressing two types of fluorescent ubiquitination-based cell cycle indicators (Fucci), HeLa-Fucci (CA)2 and HeLa-Fucci (SA), and combined this approach with the micronucleus (MN) assay to assess radioresponse. The Fucci system distinguishes cell cycle phases based on the colour of fluorescence and cell morphology under live conditions. Time-lapse imaging allowed us to further identify sub-positions within the G1 and S phases at the time of irradiation by two independent means, and to quantitate the number of MNs by following each cell through M phase until the next G1 phase. Notably, we found that radioresponse was low in late G1 phase, but rapidly increased in early S phase. It then decreased until late S phase and increased in G2 phase. For the first time, we demonstrated the unique fluctuation of radioresponse by the MN assay during the cell cycle in HeLa cells. We discuss the difference between previous clonogenic experiments using M phase-synchronised cell populations and ours, as well as the clinical implications of the present findings.

scholarly journals 72 EFFECT OF CELL CYCLE PHASE OF GENE-MANIPULATED FETAL FIBROBLASTS ON THE DEVELOPMENT OF CLONED BOVINE EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 186
Author(s):  
M. Urakawa ◽  
T. Sawada ◽  
Y. Sendai ◽  
Y. Shinkai ◽  
A. Ideta ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Donor Cell ◽  
Developmental Rate ◽  
G1 Phase ◽  
Control Group ◽  
Rate Of Development ◽  
Normal Fetus ◽  
Fetal Fibroblasts ◽  
Blastocyst Rate ◽  
And Control

Transgenic bovine fetuses and offspring can be produced by using gene-modified somatic cells and clones of these cells. In this study, we examined the effects of specific cell cycle (early G1 phase) of donor cell (gene-manipulated fibroblasts) on the development of the nuclear transfer (NT) embryos into blastocysts and on the fetus production after embryo transfer. The gene-manipulated (tg; targeting of one or both alleles of gene encoding α-1,3-galactosyltransferase) or non-manipulated (control) bovine fetal fibroblasts were used for NT. The fibroblasts transfected with the targeting vector were selected with 0.4 mg mL−1 G418. The G418-resistant cells were monitored by PCR and Southern blot analysis. The cells (tg cells) in which homologous recombination occurred were used for NT. For NT, both tg cells and control cells were cultured in DMEM with 10% FCS. Early G1 cells were prepared by choosing pairs of bridged cells derived from mitotic phase cells (Urakawa M et al. 2004 Theriogenology 62, 714–728), and non-synchronized cells were obtained from a culture plate that had reached 60–80% confluence. Each donor cell was inserted into an enucleated, in vitro-matured (19 h) oocyte. Oocyte-cell couples were electrofused and activated with calcium ionophore and cycloheximide. The NT embryos were then co-cultured with bovine oviduct epithelial cells in CR1aa with 5% CS. The blastocyst rates were determined at 6 days after NT. The blastocysts were nonsurgically transferred to recipient heifers, and the developmental rate to the normal fetus was examined by the recovery of fetus or by using ultrasonography at Days 35–42. Data were analyzed by ANOVA. The developmental rate to the blastocyst stage did not differ significantly between tg (28.4%, 128/425) and control (25.4%, 181/739) cell groups. In the control group, the blastocyst rate of embryos constructed from early G1 phase fibroblasts (25.7%, 80/311) was not significantly different from that of embryos constructed with non-synchronized fibroblasts (23.6%, 101/428). In contrast, the blastocyst rate of tg cell derived-embryos was lower (P < 0.05) in early G1 phase (23.5%, 71/302) than in non-synchronized cell phase (46.3%, 57/123). The rate of development to a normal fetus in the tg group (15.4%, 4/26) was significant lower than that in the control group (62.5%, 25/40). For both the tg group and the control group, the rate of development to fetus tended to be higher (P > 0.05) for blastocysts derived from cells at the early G1 phase than for blastocysts derived from non-synchronized cells (tg group, 25.0%, 3/12 v. 7.1%, 1/14; control group, 90.0%, 9/10 v. 53.3%, 16/30). These results demonstrate that gene modification of fetal fibroblasts affects the development of NT embryos to fetuses. In addition, the synchronization of genetically modified donor cells to the early G1 phase may increase the potential to develop to a normal fetus.

scholarly journals Estrogen Receptor Beta Displays Cell Cycle-Dependent Expression and Regulates the G1 Phase through a Non-Genomic Mechanism in Prostate Carcinoma Cells

2008 ◽  
Vol 30 (4) ◽  
pp. 349-365 ◽  
Author(s):  
Antoni Hurtado ◽  
Tomàs Pinós ◽  
Anna Barbosa-Desongles ◽  
Sandra López-Avilés ◽  
Jordi Barquinero ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Estrogen Receptor ◽  
Estrogen Receptor Beta ◽  
Mechanisms Of Action ◽  
S Phase ◽  
G1 Phase ◽  
Over Expression ◽  
M Phase ◽  
Receptor Beta

Background: It is well known that estrogens regulate cell cycle progression, but the specific contributions and mechanisms of action of the estrogen receptor beta (ERβ) remain elusive.Methods: We have analyzed the levels of ERβ1 and ERβ2 throughout the cell cycle, as well as the mechanisms of action and the consequences of the over-expression of ERβ1 in the human prostate cancer LNCaP cell line.Results: Both ERβ1 mRNA and protein expression increased from the G1 to the S phase and decreased before entering the G2/M phase, whereas ERβ2 levels decreased during the S phase and increased in the G2/M phase. ERβ1 protein was detected in both the nuclear and non-nuclear fractions, and ERβ2 was found exclusively in the nucleus. Regarding the mechanisms of action, endogenous ERβ was able to activate transcription via ERE during the S phase in a ligand-dependent manner, whereas no changes in AP1 and NFκB transactivation were observed after exposure to estradiol or the specific inhibitor ICI 182,780. Over-expression of either wild type ERβ1 or ERβ1 mutated in the DNA-binding domain caused an arrest in early G1. This arrest was accompanied by the interaction of over-expressed ERβ1 with c-Jun N-terminal protein kinase 1 (JNK1) and a decrease in c-Jun phosphorylation and cyclin D1 expression. The administration of ICI impeded the JNK1–ERβ1 interaction, increased c-Jun phosphorylation and cyclin D1 expression and allowed the cells to progress to late G1, where they became arrested.Conclusions: Our results demonstrate that, in LNCaP prostate cancer cells, both ERβ isoforms are differentially expressed during the cell cycle and that ERβ regulates the G1 phase by a non-genomic mechanism.

scholarly journals Evaluating the effects of simulated microgravity on mouse fibroblast

2020 ◽  
Vol 42 (4) ◽  
Author(s):  
Hoang Nghia Son ◽  
Hoang Nguyen Quang Huy ◽  
Tran Thi Bich Tram ◽  
Ly Ngoc Cang ◽  
Ho Nguyen Quynh Chi ◽  
...  
Keyword(s):  
Protein Expression ◽  
Simulated Microgravity ◽  
Mouse Fibroblast ◽  
S Phase ◽  
G1 Phase ◽  
Cytoskeletal Protein ◽  
Control Group ◽  
Nuclear Area ◽  
Mouse Fibroblasts ◽  
M Phase

The present study investigated how mouse fibroblasts changed under microgravity (SMG) conditions (< 10-3 G) simulated by 3D clinostat. Results showed that SMG condition markedly reduced the proliferation of mouse fibroblasts, significantly reducing the nuclear area and intensity. Compared to the control group, the mouse fibroblasts ratio of the SMG group was higher in the G0/G1 phase but lower in the S phase and G2/M phase. The ratios of early and late apoptotic cells were also higher in the SMG group. The mouse fibroblasts under SMG conditions exhibited a reduction of β-Actin and α-Tubulin 3 expressions compared to the control group. These results suggested that the SMG condition diminished the proliferation and downregulated cytoskeletal protein expression of mouse fibroblasts. 

The effects of gemcitabine on cell apoptosis and cycle of gastric cancer

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 15181-15181
Author(s):  
L. Wang
Keyword(s):  
Gastric Cancer ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Apoptosis ◽  
Morphological Changes ◽  
S Phase ◽  
Control Group ◽  
Control Groups ◽  
Gastric Cancer Cells ◽  
M Phase

15181 Background: To study the effects of gemcitabine on cell apoptosis and cell cycle of gastric cancer Methods: Gastric cancer cells were cultured with different concentrations of gemcitabine (0.001, 0.01 and 0.1μM). MTT test was performed to evaluate the cell proliferation. The cells were divided into three groups: control group (cultured in RPMI-1640) and 5-FU group ( cultured in RPMI-1640 with 5- FU) and gemcitabine group ( cultured in RPMI-1640 with Gemcitabine). Flow cytometry was performed to determine the apoptotic rate and the cell cycle phases. Morphological changes were observed by phasecontrast microscope. Results: The cell proliferation was inhibited in experiment groups treated with gemcitabine and 5-FU, compared with control groups(P<0.01). Gemcitabine can induce cell apoptosis. 0.01μM and 0.1μM gemcitabine were much more effective than 0.001μM. On the third day, S phase cells accounted for 24.5% and G2-M phase cells 0.08% in the control group, while 24.6% and 0.06%, respectively in the gemcitabine group. However, on the seventh day, those came to 20.8% and 0.41% in the control group, and 18.2% and 1. 55% in the gemcitabine group, indicating a significant change in the cell cycle ( P<0.01). Conclusions: Gemcitabine can inhibit the cell proliferation, and it maybe related to cell apoptosis. No significant financial relationships to disclose.

scholarly journals Novel phase I study combining G1 phase, S phase, and G2/M phase cell cycle inhibitors in patients with advanced malignancies

Cell Cycle ◽  
2015 ◽  
Vol 14 (21) ◽  
pp. 3434-3440
Author(s):  
Rajul K Jain ◽  
David S Hong ◽  
Aung Naing ◽  
Jennifer Wheler ◽  
Thorunn Helgason ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Phase I ◽  
Phase I Study ◽  
S Phase ◽  
Phase Cell ◽  
G1 Phase ◽  
Advanced Malignancies ◽  
M Phase ◽  
Cell Cycle Inhibitors

Cell-cycle blockage associated with increased apoptotic cells in the thymus of chickens fed on diets high in fluorine

2010 ◽  
Vol 30 (7) ◽  
pp. 685-692 ◽  
Author(s):  
Tao Chen ◽  
Hengmin Cui ◽  
Yun Cui ◽  
Caimin Bai ◽  
Tao Gong ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Control Diet ◽  
Growth Index ◽  
S Phase ◽  
Control Group ◽  
Group Iii ◽  
Group I ◽  
M Phase ◽  
Cellular Apoptosis ◽  
Cellular Immune

Three hundred 1-day-old Avian broilers were divided into four groups and fed on control diet (fluorine 23 mg/kg) and high-fluorine (F) diets (400 mg/kg, high-F group I; 800 mg/kg, high-F group II; 1200 mg/kg, high-F group III) for 42 days (n = 75/group). The growth index (GI) was obviously decreased in the three high-F groups, which indicated the inhibited development of thymus. Histopathologically, the population of thymocytes was decreased in the thymic lobule in the three high-F groups. As measured by flow cytometry, thymocytes in G0/G 1 phase were significantly increased while thymocytes in S phase, G 2 + M phase and proliferating index (PI) value were obviously decreased in the three high-F groups. Also, the percentage of apoptotic thymocytes was greatly increased in the three high-F groups when compared with that of control group. At the same time, the occurrence frequencies of apoptotic thymocyte were markedly increased in the three high-F groups, with the appearance of dilated endoplasmic reticulum in high-F groups II and III ultra-structurally. The results showed that excess dietary F in the range of 400-1200 mg/kg caused histological lesions, G0/G1 arrest and cellular apoptosis in the thymus, which inhibited the development of thymus and finally led to impaired cellular immune function.

scholarly journals The Paramyxovirus Simian Virus 5 V Protein Slows Progression of the Cell Cycle

2000 ◽  
Vol 74 (19) ◽  
pp. 9152-9166 ◽  
Author(s):  
Grace Y. Lin ◽  
Robert A. Lamb
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Simian Virus ◽  
S Phase ◽  
V Protein ◽  
Simian Virus 5 ◽  
C Terminus ◽  
M Phase ◽  
Infected Cells ◽  
Affect Cell Cycle Progression

ABSTRACT Infection of cells by many viruses affects the cell division cycle of the host cell to favor viral replication. We examined the ability of the paramyxovirus simian parainfluenza virus 5 (SV5) to affect cell cycle progression, and we found that SV5 slows the rate of proliferation of HeLa T4 cells. The SV5-infected cells had a delayed transition from G1 to S phase and prolonged progression through S phase, and some of the infected cells were arrested in G2 or M phase. The levels of p53 and p21CIP1were not increased in SV5-infected cells compared to mock-infected cells, suggesting that the changes in the cell cycle occur through a p53-independent mechanism. However, the phosphorylation of the retinoblastoma protein (pRB) was delayed and prolonged in SV5-infected cells. The changes in the cell cycle were also observed in cells expressing the SV5 V protein but not in the cells expressing the SV5 P protein or the V protein lacking its unique C terminus (VΔC). The unique C terminus of the V protein of SV5 was shown previously to interact with DDB1, which is the 127-kDa subunit of the multifunctional damage-specific DNA-binding protein (DDB) heterodimer. The coexpression of DDB1 with V can partially restore the changes in the cell cycle caused by expression of the V protein.

scholarly journals CLN3 expression is sufficient to restore G1-to-S-phase progression in Saccharomyces cerevisiae mutants defective in translation initiation factor eIF4E

1999 ◽  
Vol 340 (1) ◽  
pp. 135-141 ◽  
Author(s):  
Parisa DANAIE ◽  
Michael ALTMANN ◽  
Michael N. HALL ◽  
Hans TRACHSEL ◽  
Stephen B. HELLIWELL
Keyword(s):  
Cell Cycle ◽  
S Phase ◽  
Translation Initiation Factor ◽  
Yeast Cells ◽  
Initiation Factor ◽  
G1 Phase ◽  
Mrna Levels ◽  
Wild Type ◽  
Phase Progression ◽  
Type Gene

The essential cap-binding protein (eIF4E) of Saccharomycescerevisiae is encoded by the CDC33 (wild-type) gene, originally isolated as a mutant, cdc33-1, which arrests growth in the G1 phase of the cell cycle at 37 °C. We show that other cdc33 mutants also arrest in G1. One of the first events required for G1-to-S-phase progression is the increased expression of cyclin 3. Constructs carrying the 5ʹ-untranslated region of CLN3 fused to lacZ exhibit weak reporter activity, which is significantly decreased in a cdc33-1 mutant, implying that CLN3 mRNA is an inefficiently translated mRNA that is sensitive to perturbations in the translation machinery. A cdc33-1 strain expressing either stable Cln3p (Cln3-1p) or a hybrid UBI4 5ʹ-CLN3 mRNA, whose translation displays decreased dependence on eIF4E, arrested randomly in the cell cycle. In these cells CLN2 mRNA levels remained high, indicating that Cln3p activity is maintained. Induction of a hybrid UBI4 5ʹ-CLN3 message in a cdc33-1 mutant previously arrested in G1 also caused entry into a new cell cycle. We conclude that eIF4E activity in the G1-phase is critical in allowing sufficient Cln3p activity to enable yeast cells to enter a new cell cycle.

scholarly journals Posttranslational Phosphorylation and Ubiquitination of the Saccharomyces cerevisiae Poly(A) Polymerase at the S/G2 Stage of the Cell Cycle

2000 ◽  
Vol 20 (8) ◽  
pp. 2794-2802 ◽  
Author(s):  
Neptune Mizrahi ◽  
Claire Moore
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Sorting ◽  
S Phase ◽  
Phase Arrest ◽  
Phosphatase Treatment ◽  
M Phase ◽  
64 Kda Protein ◽  
Mitotic Phosphorylation ◽  
Cycle Regulation

ABSTRACT The poly(A) polymerase of the budding yeast Saccharomyces cerevisiae (Pap1) is a 64-kDa protein essential for the maturation of mRNA. We have found that a modified Pap1 of 90 kDa transiently appears in cells after release from α-factor-induced G1 arrest or from a hydroxyurea-induced S-phase arrest. While a small amount of modification occurs in hydroxyurea-arrested cells, fluorescence-activated cell sorting analysis and microscopic examination of bud formation indicate that the majority of modified enzyme is found at late S/G2 and disappears by the time cells have reached M phase. The reduction of the 90-kDa product upon phosphatase treatment indicates that the altered mobility is due to phosphorylation. A preparation containing primarily the phosphorylated Pap1 has no poly(A) addition activity, but this activity is restored by phosphatase treatment. A portion of Pap1 is also polyubiquitinated concurrent with phosphorylation. However, the bulk of the 64-kDa Pap1 is a stable protein with a half-life of 14 h. The timing, nature, and extent of Pap1 modification in comparison to the mitotic phosphorylation of mammalian poly(A) polymerase suggest an intriguing difference in the cell cycle regulation of this enzyme in yeast and mammalian systems.

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