scholarly journals Decomposition of cell activities revealing the role of the cell cycle in driving biofunctional heterogeneity

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Tian Lan ◽  
Meng Yu ◽  
Weisheng Chen ◽  
Jun Yin ◽  
Hsiang-Tsun Chang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Drug Resistance ◽  
Cell Migration ◽  
Cell Morphology ◽  
Rho Gtpases ◽  
Kinase Inhibitors ◽  
Cell Phenotype ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
The Relationship

AbstractHeterogeneity of cell phenotypes remains a barrier in progressing cell research and a challenge in conquering cancer-related drug resistance. Cell morphology, the most direct property of cell phenotype, evolves along the progression of the cell cycle; meanwhile, cell motility, the dynamic property of cell phenotype, also alters over the cell cycle. However, a quantifiable research understanding the relationship between the cell cycle and cell migration is missing. Herein, we coordinate the migratory behaviours of NIH 3T3 fibroblasts to their corresponding phases of the cell cycle, the G1, the S, and the G2 phases, and explain the relationship through the spatiotemporal arrangements between the Rho GTPases’ signals and cyclin-dependent kinase inhibitors, p21Cip1, and p27Kip1. Taken together, we demonstrate that both cell morphology and the dynamic subcellular behaviour are homogenous within each stage of the cell cycle phases but heterogenous between phases through quantitative cell analyses and an interactive molecular mechanism between the cell cycle and cell migration, posing potential implications in countering drug resistance.

scholarly journals Phenotype Spectrum reflects Synergies among the Cell Architecture over Stages of the Cell Cycle

2020 ◽  
Author(s):  
Tian Lan ◽  
Meng Yu ◽  
Weisheng Chen ◽  
Jun Yin ◽  
Hsiang-Tsun Chang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Drug Resistance ◽  
Cell Migration ◽  
Cell Morphology ◽  
Rho Gtpases ◽  
Kinase Inhibitors ◽  
Cell Phenotype ◽  
And Migration ◽  
Cell Phenotypes ◽  
Nih 3T3

AbstractThe heterogeneity of cell phenotypes remains a barrier in progressing cell research and a challenge in conquering cancer-related drug resistance. Cell morphology, the most direct property of cell phenotype, evolves along the progression of the cell cycle; meanwhile, cell motility, the dynamic property of cell phenotype, also alters over the cell cycle. However, a quantifiable research understanding the strict relationship between the cell cycle and cell migration is missing. Herein, we separately elucidate the correspondence of single NIH 3T3 fibroblast migratory behaviors with the G1, S, and G2 phases of the cell cycle, an underlying property of proliferation. The results show that synergies among the highly spatiotemporal arrangements of signals in Rho GTPases and cyclin-dependent kinase inhibitors, p21Cip1, and p27Kip1 coordinates proliferation and migration. Taken together, we explain the synergies among these processes through providing an interactive molecular mechanism between the cell cycle and cell migration and demonstrate that both cell morphology and the dynamic subcellular behavior are homogenous within each stage of the cell cycle phases, posing potential implications in countering drug resistance.

scholarly journals In Vivo Fluconazole Pharmacodynamics and Resistance Development in a Previously Susceptible Candida albicans Population Examined by Microbiologic and Transcriptional Profiling

2006 ◽  
Vol 50 (7) ◽  
pp. 2384-2394 ◽  
Author(s):  
D. Andes ◽  
A. Lepak ◽  
J. Nett ◽  
L. Lincoln ◽  
K. Marchillo
Keyword(s):  
Drug Resistance ◽  
Candida Albicans ◽  
Transcriptional Profiling ◽  
Cell Phenotype ◽  
Cell Populations ◽  
Dosing Regimen ◽  
Resistance Development ◽  
The Impact ◽  
The Relationship

ABSTRACT Antimicrobial drug resistance can limit the ability to effectively treat patients. Numerous factors have been proposed to impact the development of antimicrobial resistance, including those specific to the drug and the dosing regimen. The field of investigation that examines the relationship between dosing regimen and outcome is termed antimicrobial pharmacokinetics and pharmacodynamics. Our prior in vivo investigations examined the relationship between fluconazole pharmacodynamics and the modulation of isogenic resistant and susceptible Candida albicans populations in a mixed-inoculum design (1). The goal of the current studies was to examine the impact of fluconazole pharmacodynamics on resistance emergence from a susceptible parent population over time using a murine systemic-candidiasis model. Both microbiologic and transcriptional endpoints were examined during the evolution of cell populations. As in our previous investigation, the more frequently administered dosing regimen prevented the emergence of a resistant cell phenotype. Conversely, dosing regimens that produced prolonged sub-MIC concentrations were associated with resistance development. The studies also demonstrated a striking relationship between fluconazole pharmacodynamic exposures and the mRNA abundance of drug resistance-associated efflux pumps. Global transcriptional profiling of cell populations during the progressive emergence of a resistance phenotype provides insight into the mechanisms underlying this complex physiologic process.

scholarly journals A cell cycle and mutational analysis of anchorage-independent growth: cell adhesion and TGF-beta 1 control G1/S transit specifically

1993 ◽  
Vol 122 (2) ◽  
pp. 461-471 ◽  
Author(s):  
EK Han ◽  
TM Guadagno ◽  
SL Dalton ◽  
RK Assoian
Keyword(s):  
Cell Cycle ◽  
Growth Factor ◽  
Cell Cycle Progression ◽  
Fibroblast Cell ◽  
Tgf Beta ◽  
Cycle Progression ◽  
Anchorage Independent Growth ◽  
Differential Effects ◽  
3T3 Fibroblasts ◽  
Nih 3T3

We have examined cell cycle control of anchorage-independent growth in nontransformed fibroblasts. In previous studies using G0-synchronized NRK and NIH-3T3 cells, we showed that anchorage-independent growth is regulated by an attachment-dependent transition at G1/S that resembles the START control point in the cell cycle of Saccharomyces cerevisiae. In the studies reported here, we have synchronized NRK and NIH-3T3 fibroblasts immediately after this attachment-dependent transition to determine if other portions of the fibroblast cell cycle are similarly regulated by adhesion. Our results show that S-, G2-, and M-phase progression proceed in the absence of attachment. Thus, we conclude that the adhesion requirement for proliferation of these cells can be explained in terms of the single START-like transition. In related studies, we show that TGF-beta 1 overrides the attachment-dependent transition in NRK and AKR-2B fibroblasts (lines in which TGF-beta 1 induces anchorage-independent growth), but not in NIH-3T3 or Balb/c 3T3 fibroblasts (lines in which TGF-beta 1 fails to induce anchorage-independent growth). These results show that (a) adhesion and TGF-beta 1 can have similar effects in stimulating cell cycle progression from G1 to S and (b) the differential effects of TGF-beta 1 on anchorage-independent growth of various fibroblast lines are directly reflected in the differential effects of the growth factor at G1/S. Finally, we have randomly mutagenized NRK fibroblasts to generate mutant lines that have lost their attachment/TGF-beta 1 requirement for G1/S transit while retaining their normal mitogen requirements for proliferation. These clones, which readily proliferate in mitogen-supplemented soft agar, appear non-transformed in monolayer: they are well spread, nonrefractile, and contact inhibited. The existence of this new fibroblast phenotype demonstrates (a) that the growth factor and adhesion/TGF-beta 1 requirements for cell cycle progression are genetically separable, (b) that the two major control points in the fibroblast cell cycle (G0/G1 and G1/S) are regulated by distinct extracellular signals, and (c) that the genes regulating anchorage-independent growth need not be involved in regulating contact inhibition, focus formation, or growth factor dependence.

scholarly journals Ethanol, Zn2+ and insulin interact as progression factors to enhance DNA synthesis synergistically in the presence of Ca2+ and other cell cycle initiators in fibroblasts

2000 ◽  
Vol 346 (1) ◽  
pp. 241-247 ◽  
Author(s):  
Jin-Sheng HUANG ◽  
Qing-Bai SHE ◽  
Karan S. CRILLY ◽  
Zoltan KISS
Keyword(s):  
Cell Cycle ◽  
Growth Factor ◽  
Dna Synthesis ◽  
Cell Cycle Progression ◽  
Synergistic Effects ◽  
Dependent Manner ◽  
Cycle Progression ◽  
3T3 Fibroblasts ◽  
Igf I ◽  
Nih 3T3

In serum-starved NIH 3T3 fibroblasts, ethanol (30-80 mM) promoted the effects of insulin and insulin-like growth factor I (IGF-I) on DNA synthesis in a Zn2+-dependent manner. Ethanol and Zn2+ were most effective when added shortly before or after insulin, indicating that all these agents facilitated cell cycle progression. The synergistic effects of ethanol, Zn2+ and insulin (or IGF-I) on DNA synthesis required 1.1-2.3 mM Ca2+, which seemed to act as the cell cycle initiator. When serum-starved cells were pretreated for 2 h with other cell cycle initiators such as 10% (v/v) serum, 50 ng/ml platelet-derived growth factor or 2 ng/ml fibroblast growth factor, subsequent co-treatments with 60 mM ethanol, Zn2+ and insulin for an 18 h period again synergistically increased DNA synthesis. Of the various signal transducing events examined, ethanol stimulated cellular uptake of 45Ca and it enhanced the stimulatory effects of insulin on p70 S6 kinase activity in a Zn2+-dependent manner. In contrast, ethanol inhibited insulin-induced activating phosphorylation of p42/p44 mitogen-activated protein kinases; these inhibitory ethanol effects were prevented by Zn2+. The results show that, in NIH 3T3 fibroblasts, ethanol can promote cell cycle progression in the presence of a cell cycle initiator as well as Zn2+ and insulin (or IGF-I).

scholarly journals 5-Methoxyhydnocarpin shows selective anti-cancer effects and induces apoptosis in THP-1 human leukemia cancer cells via mitochondrial disruption, suppression of cell migration and invasion and cell cycle arrest

2016 ◽  
Vol 11 (3) ◽  
pp. 652
Author(s):  
Xiao-Yan Xu ◽  
Hui-Rui Wang ◽  
Shu-Li Guo ◽  
Bo Li ◽  
Tao Lv
Keyword(s):  
Cell Cycle ◽  
Cell Migration ◽  
Cell Cycle Arrest ◽  
Cell Morphology ◽  
Phase Distribution ◽  
Migration And Invasion ◽  
Cycle Phase ◽  
Human Leukemia ◽  
Cytotoxic Effects ◽  
Cycle Arrest

<p class="Abstract">The primary objective of the current investigation was to study the anti-tumor effects of 5-methoxyhydnocarpin in THP-1 human leukemia cells along with evaluating its effects on apoptotic induction, cancer cell migration and cell cycle phase distribution. MTT cell viability assay revealed the cell cytotoxic effects while as clonogenic assay revealed the effects of the compound on colony formation tendency. Phase contrast microscopy revealed the effects on cell morphology while as its effects on cell cycle and apoptosis induction were evaluated by flow cytometer. Results showed that 5-methoxyhydnocarpin induced both anchorage-dependent and anchorage-independent cytotoxic effects in THP-1 cells. 5-Methoxyhydnocarpin-treated cells exhibited significant changes in cell morphology. Flow cytometry experiment indicated that 5-methoxyhydnocarpin led to sub-G1 cell cycle arrest and also led to early and late apoptosis.</p><p class="Abstract"><strong>Video Clips:</strong></p><p class="Abstract"><a href="https://www.youtube.com/v/GXx829BOW0U">1</a>  <a href="https://www.youtube.com/v/FaN-UcPk2vo">2</a>  <a href="https://www.youtube.com/v/V1Qp-Ibj3-8">3</a>  <a href="https://www.youtube.com/v/crqEjgvgFE0">4</a>  <a href="https://www.youtube.com/v/r7QXXlaqzCk">5</a>  <a href="https://www.youtube.com/v/unGjmzO9Nhk">6</a>  <a href="https://www.youtube.com/v/RpBydoaSji8">7</a> </p><p class="Abstract"> </p><p> </p>

scholarly journals c-Ha-rasVal 12 oncogene-transformed NIH-3T3 fibroblasts display more decondensed nucleosomal organization than normal fibroblasts.

1990 ◽  
Vol 111 (1) ◽  
pp. 9-17 ◽  
Author(s):  
J Laitinen ◽  
L Sistonen ◽  
K Alitalo ◽  
E Hölttä
Keyword(s):  
Cell Cycle ◽  
Micrococcal Nuclease ◽  
Normal Fibroblast ◽  
Transformed Cells ◽  
Serum Starvation ◽  
Quiescent State ◽  
Normal Cells ◽  
3T3 Fibroblasts ◽  
Mnase Digestion ◽  
Nih 3T3

We have compared the nucleosomal organization of c-Ha-rasVal 12 oncogene-transformed NIH-3T3 fibroblasts with that of normal fibroblasts by using micrococcal nuclease (MNase) as a probe for the chromatin structure. The bulk chromatin from asynchronously and exponentially growing ras-transformed cells was much more sensitive to MNase digestion than chromatin from the normal cells. Southern hybridization analyses of the MNase digests with probes specific for the ornithine decarboxylase (odc) and c-myc genes showed that the coding and/or 3' end regions of these growth-inducible genes carry a nucleosomal organization both in ras-transformed and normal cells. Studies with cells synchronized by serum starvation showed that in both cell lines the nucleosomal organization of chromatin is relatively condensed at the quiescent state, becomes highly decondensed during the late G1 phase of the cell cycle, and starts again to condense during the S phase. However, in ras-transformed cells the decondensation state stayed much longer than in normal cells. Moreover, irrespective of the phase of the cell cycle the bulk chromatin as well as that of the odc and c-myc genes was more sensitive to MNase digestion in the ras-transformed cell than in the normal fibroblast. Decondensation of the chromatin was also observed in the normal c-Ha-ras protooncogene-transfected cells, but to a lesser extent than in the mutant ras-transformed cells. Whether the increased degree of chromatin decondensation plays a regulatory role in the increased expression of many growth-related genes in the ras-transformed cells remains an interesting object of further study.

scholarly journals Rac Regulates Endothelial Morphogenesis and Capillary Assembly

2002 ◽  
Vol 13 (7) ◽  
pp. 2474-2485 ◽  
Author(s):  
John O. Connolly ◽  
Nandi Simpson ◽  
Lindsay Hewlett ◽  
Alan Hall
Keyword(s):  
Cell Migration ◽  
Endothelial Cell ◽  
Cell Morphology ◽  
Rho Gtpases ◽  
Actin Polymerization ◽  
Capillary Tube ◽  
Branching Morphogenesis ◽  
Small Gtpase ◽  
Capillary Tubes

Endothelial cells undergo branching morphogenesis to form capillary tubes. We have utilized an in vitro Matrigel overlay assay to analyze the role of the cytoskeleton and Rho GTPases during this process. The addition of matrix first induces changes in cell morphology characterized by the formation of dynamic cellular protrusions and the assembly of discrete aggregates or cords of aligned cells resembling primitive capillary-like structures, but without a recognizable lumen. This is followed by cell migration leading to the formation of a complex interconnecting network of capillary tubes with readily identifiable lumens. Inhibition of actin polymerization or actin-myosin contraction inhibits cell migration but has no effect on the initial changes in endothelial cell morphology. However, inhibition of microtubule dynamics prevents both the initial cell shape changes as well as cell migration. We find that the small GTPase Rac is essential for the matrix-induced changes in endothelial cell morphology, whereas p21-activated kinase, an effector of Rac, is required for cell motility. We conclude that Rac integrates signaling through both the actin and microtubule cytoskeletons to promote capillary tube assembly.

scholarly journals Inositol polyphosphates are not increased by overexpression of Ins(1,4,5)P3 3-kinase but show cell-cycle dependent changes in growth factor-stimulated fibroblasts.

1994 ◽  
Vol 5 (1) ◽  
pp. 17-27 ◽  
Author(s):  
T Balla ◽  
S S Sim ◽  
A J Baukal ◽  
S G Rhee ◽  
K J Catt
Keyword(s):  
Cell Cycle ◽  
Growth Factor ◽  
Inositol Polyphosphates ◽  
Transfected Cells ◽  
3T3 Fibroblasts ◽  
Significant Difference ◽  
Early Peak ◽  
Tightly Coupled ◽  
Cycle Dependent ◽  
Nih 3T3

NIH 3T3 fibroblasts were stably transfected with rat brain inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) 3-kinase to explore the relationship between increased production of Ins(1,3,4,5)P4 and the formation of InsP5 and InsP6. Mass measurements of InsP5 and InsP6 revealed no significant difference between kinase- and vector-transfected fibroblasts. However, such 3-kinase-transfected cells, when labeled with [3H]inositol for 48-72 h, showed lower levels of [3H]InsP5 and [3H]InsP6, as well as [3H]Ins(1,3,4,6)P4 and D/L[3H]Ins(1,4,5,6)P4, than their vector-transfected counterparts. Because Ins(1,4,5)P3 3-kinase-transfected cells grew less rapidly than vector-transfected controls, we determined whether the synthesis of InsP5 and InsP6 was related to a specific phase of the cell cycle. When NIH 3T3 cells prelabeled with [3H]inositol were synchronized by serum deprivation followed by stimulation with platelet-derived growth factor (PDGF), the amounts of labeled InsP5 and InsP6 began to increase only after 12 h of stimulation, when cells entered the S-phase as indicated by increased [3H]thymidine incorporation. The enhanced synthesis of these inositol polyphosphates was preceded by an early increase in Ins(1,4,5)P3 and its metabolites that was no longer evident by the fifth hour of PDGF action. There was also a prominent and biphasic increase in the level of D/L-Ins(1,4,5,6)P4 with an early peak at approximately 3 h and a second rise that paralleled the increases in InsP5 and InsP6. These results indicate that the formation of highly phosphorylated inositols is not tightly coupled to the receptor-mediated formation of Ins(1,4,5)P3 and its metabolites but is mainly determined by other factors that operate at specific points of the cell cycle.

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