Cell cycle specific expression of “tissue” transglutaminase (tTG) in apoptotic cells

Rationale: The regenerative capacity of the heart to repair itself after myocardial infarction (MI)is limited. Our previous study showed that ectopic introduction of Cdk1/CyclinB1 andCdk4/CyclinD1 complexes (4F) promotes cardiomyocyte proliferation in vitro and in vivo andimproves cardiac function after MI. However, its clinical application is limited due to the concernsfor tumorigenic potential in other organs. Objectives: To first, identify on a single cell transcriptomic basis the necessary reprogrammingsteps that cardiomyocytes need to undertake to progress through the proliferation processfollowing 4F overexpression, and then, to determine the pre-clinical efficacy of transient andcardiomyocyte specific expression of 4F in improving cardiac function after MI in small and largeanimals. Methods and Results: Temporal bulk and single cell RNAseq of mature hiPS-CMs treated with4F or LacZ control for 24, 48, or 72 h revealed full cell cycle reprogramming in 15% of thecardiomyocyte population which was associated with sarcomere disassembly and metabolicreprogramming. Transient overexpression of 4F specifically in cardiomyocytes was achievedusing non-integrating lentivirus (NIL) driven by TNNT2 (TNNT2-4F-NIL). One week after inductionof ischemia-reperfusion injury in rats or pigs, TNNT2-4F-NIL or control virus was injectedintramyocardially. Compared with controls, rats or pigs treated with TNNT2-4F-NIL showed a 20-30% significant improvement in ejection fraction and scar size four weeks after treatment, asassessed by echocardiography and histological analysis. Quantification of cardiomyocyteproliferation in pigs using a novel cytokinesis reporter showed that ~10% of the cardiomyocyteswithin the injection site were labelled as daughter cells following injection with TNNT2-4F-NILcompared with ~0.5% background labelling in control groups. Conclusions: We provide the first understanding of the process of forced cardiomyocyteproliferation and advanced the clinical applicability of this approach through minimization ofoncogenic potential of the cell cycle factors using a novel transient and cardiomyocyte-specificviral construct.

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Pharmacokinetic, Antiproliferative and Apoptotic Effects of Phenolic Acids in Human Colon Adenocarcinoma Cells Using In Vitro and In Silico Approaches

Molecules ◽

10.3390/molecules23102569 ◽

2018 ◽

Vol 23 (10) ◽

pp. 2569 ◽

Cited By ~ 13

Author(s):

Lana Rosa ◽

Nathállia Jordão ◽

Nathália da Costa Pereira Soares ◽

Joelma deMesquita ◽

Mariana Monteiro ◽

...

Keyword(s):

Cell Cycle ◽

Colon Cancer ◽

Cell Viability ◽

Colon Adenocarcinoma ◽

Human Colon ◽

G1 Phase ◽

Apoptotic Cells ◽

Adenocarcinoma Cells ◽

Human Colon Adenocarcinoma ◽

Colon Adenocarcinoma Cells

Colon cancer is the second most common cause of cancer deaths in the USA and Europe. Despite aggressive therapies, many tumors are resistant to current treatment protocols and epidemiological data suggest that diet is a major factor in the etiology of colon cancer. This study aimed to evaluate the antioxidant activity and the influence of 3,4-dihydroxyphenylacetic (3,4-DHPAA), p-coumaric (p-CoA), vanillic (VA) and ferulic (FA) acids on cell viability, cell cycle progression, and rate of apoptosis in human colon adenocarcinoma cells (HT-29). The results showed that all compounds tested reduce cell viability in human colon cancer cells. 3,4-DHPAA promoted the highest effect antiproliferative with an increase in the percentage of cells in G0/G1 phase, accompanied by a reduction of cells in G2/M phase. Cell cycle analysis of VA and FA showed a decrease in the proportion of cells in G0/G1 phase (10.0 µM and 100.0 µM). p-CoA and FA acids increased the percentage of apoptotic cells and non-apoptotic cells. 3,4-DHPAA seems to be the substance with the greatest potential for in vivo studies, opening thus a series of perspectives on the use of these compounds in the prevention and treatment of colon cancer.

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Cell cycle kinetics, tissue transglutaminase and programmed cell death (apoptosis)

FEBS Letters ◽

10.1016/0014-5793(92)81392-y ◽

1992 ◽

Vol 311 (2) ◽

pp. 174-178 ◽

Cited By ~ 29

Author(s):

S. El Alaoui ◽

S. Mian ◽

J. Lawry ◽

G. Quash ◽

M. Griffin

Keyword(s):

Cell Cycle ◽

Cell Death ◽

Programmed Cell Death ◽

Tissue Transglutaminase ◽

Cell Cycle Kinetics

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P0007 PP TISSUE TRANSGLUTAMINASE AUTOANTIBODIES REGULATE ACTIN REARRANGEMENTS AND CELL CYCLE IN SEVERAL CELL TYPES

Journal of Pediatric Gastroenterology and Nutrition ◽

10.1097/00005176-200406001-00131 ◽

2004 ◽

Vol 39 (Supplement 1) ◽

pp. S58

Author(s):

R. Troncone ◽

I. Caputo ◽

M. Barone ◽

S. Auricchio ◽

C. Esposito

Keyword(s):

Cell Cycle ◽

Tissue Transglutaminase ◽

Cell Types

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Apoptotic Fragmentation of Tricellulin

International Journal of Molecular Sciences ◽

10.3390/ijms20194882 ◽

2019 ◽

Vol 20 (19) ◽

pp. 4882 ◽

Cited By ~ 3

Author(s):

Susanne Janke ◽

Sonnhild Mittag ◽

Juliane Reiche ◽

Otmar Huber

Keyword(s):

Coiled Coil ◽

Apoptotic Cells ◽

Cleavage Sites ◽

Central Importance ◽

Specific Expression ◽

Caspase Cleavage ◽

Epithelial Homeostasis ◽

Cell Layers ◽

Specific Localization ◽

Terminal Amino

Apoptotic extrusion of cells from epithelial cell layers is of central importance for epithelial homeostasis. As a prerequisite cell–cell contacts between apoptotic cells and their neighbors have to be dissociated. Tricellular tight junctions (tTJs) represent specialized structures that seal polarized epithelial cells at sites where three cells meet and are characterized by the specific expression of tricellulin and angulins. Here, we specifically addressed the fate of tricellulin in apoptotic cells. Methods: Apoptosis was induced by staurosporine or camptothecin in MDCKII and RT-112 cells. The fate of tricellulin was analyzed by Western blotting and immunofluorescence microscopy. Caspase activity was inhibited by Z-VAD-FMK or Z-DEVD-FMK. Results: Induction of apoptosis induces the degradation of tricellulin with time. Aspartate residues 487 and 441 were identified as caspase cleavage-sites in the C-terminal coiled-coil domain of human tricellulin. Fragmentation of tricellulin was inhibited in the presence of caspase inhibitors or when Asp487 or Asp441 were mutated to asparagine. Deletion of the tricellulin C-terminal amino acids prevented binding to lipolysis-stimulated lipoprotein receptor (LSR)/angulin-1 and thus should impair specific localization of tricellulin to tTJs. Conclusions: Tricellulin is a substrate of caspases and its cleavage in consequence contributes to the dissolution of tTJs during apoptosis.

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Joint changes in RNA, RNA polymerase II, and promoter activity through the cell cycle identify non-coding RNAs involved in proliferation

10.1101/2021.02.12.430890 ◽

2021 ◽

Author(s):

Siv Anita Hegre ◽

Helle Samdal ◽

Antonin Klima ◽

Endre B. Stovner ◽

Kristin G. Nørsett ◽

...

Keyword(s):

Cell Cycle ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Promoter Activity ◽

Cycle Phase ◽

Specific Expression ◽

Chip Sequencing ◽

Non Coding Rnas ◽

Cycle Dependent ◽

Rna Levels

AbstractProper regulation of the cell cycle is necessary for normal growth and development of all organisms. Conversely, altered cell cycle regulation often underlies proliferative diseases such as cancer. Long non-coding RNAs (lncRNAs) are recognized as important regulators of gene expression and are often found dysregulated in diseases, including cancers. However, identifying lncRNAs with cell cycle functions is challenging due to their often low and cell-type specific expression. We present a highly effective method that analyses changes in promoter activity, transcription, and RNA levels for identifying genes enriched for cell cycle functions. Specifically, by combining RNA sequencing with ChIP sequencing through the cell cycle of synchronized human keratinocytes, we identified 1009 genes with cell cycle-dependent expression and correlated changes in RNA polymerase II occupancy or promoter activity as measured by histone 3 lysine 4 trimethylation (H3K4me3). These genes were highly enriched for genes with known cell cycle functions and included 59 lncRNAs. We selected four of these lncRNAs – AC005682.5, RP11-132A1.4, ZFAS1, and EPB41L4A-AS1 – for further experimental validation and found that knockdown of each of the four lncRNAs affected cell cycle phase distributions and reduced proliferation in multiple cell lines. These results show that many genes with cell cycle functions have concomitant cell-cycle dependent changes in promoter activity, transcription, and RNA levels and support that our multi-omics method is well suited for identifying lncRNAs involved in the cell cycle.

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Genome-Wide Identification and Analysis of Cell Cycle Genes in Birch

Forests ◽

10.3390/f13010120 ◽

2022 ◽

Vol 13 (1) ◽

pp. 120

Author(s):

Yijie Li ◽

Song Chen ◽

Yuhang Liu ◽

Haijiao Huang

Keyword(s):

Cell Cycle ◽

Growth And Development ◽

Betula Pendula ◽

Cell Cycle Gene ◽

Expression Data ◽

Rna Seq ◽

Specific Expression ◽

Cell Cycles ◽

Cell Cycle Genes ◽

Genome Wide

Research Highlights: This study identified the cell cycle genes in birch that likely play important roles during the plant’s growth and development. This analysis provides a basis for understanding the regulatory mechanism of various cell cycles in Betula pendula Roth. Background and Objectives: The cell cycle factors not only influence cell cycles progression together, but also regulate accretion, division, and differentiation of cells, and then regulate growth and development of the plant. In this study, we identified the putative cell cycle genes in the B. pendula genome, based on the annotated cell cycle genes in Arabidopsis thaliana (L.) Heynh. It can be used as a basis for further functional research. Materials and Methods: RNA-seq technology was used to determine the transcription abundance of all cell cycle genes in xylem, roots, leaves, and floral tissues. Results: We identified 59 cell cycle gene models in the genome of B. pendula, with 17 highly expression genes among them. These genes were BpCDKA.1, BpCDKB1.1, BpCDKB2.1, BpCKS1.2, BpCYCB1.1, BpCYCB1.2, BpCYCB2.1, BpCYCD3.1, BpCYCD3.5, BpDEL1, BpDpa2, BpE2Fa, BpE2Fb, BpKRP1, BpKRP2, BpRb1, and BpWEE1. Conclusions: By combining phylogenetic analysis and tissue-specific expression data, we identified 17 core cell cycle genes in the Betulapendula genome.

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Predicting cell-cycle expressed genes identifies canonical and non-canonical regulators of time-specific expression in Saccharomyces cerevisiae

10.1101/387050 ◽

2018 ◽

Author(s):

Nicholas L Panchy ◽

John P. Lloyd ◽

Shin-Han Shiu

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Target Genes ◽

Specific Cell ◽

Data Sets ◽

Cell Cycle Regulators ◽

Specific Expression ◽

Regulatory Data ◽

Time Specific ◽

Cycle Regulation

AbstractThe collection all TFs, target genes and their interactions in an organism form a gene regulatory network (GRN), which underly complex patterns of transcription even in unicellular species. However, identifying which interactions regulate expression in a specific temporal context remains a challenging task. With multiple experimental and computational approaches to characterize GRNs, we predicted general and phase-specific cell-cycle expression in Saccharomyces cerevisiae using four regulatory data sets: chromatin immunoprecipitation (ChIP), TF deletion data (Deletion), protein binding microarrays (PBMs), and position weight matrices (PWMs). Our results indicate that the source of regulatory interaction information significantly impacts our ability to predict cell-cycle expression where the best model was constructed by combining selected TF features from ChIP and Deletion data as well as TF-TF interaction features in the form of feed-forward loops. The TFs that were the best predictors of cell-cycle expression were enriched for known cell-cycle regulators but also include regulators not implicated in cell-cycle regulation previously. In addition, ChIP and Deletion datasets led to the identification different subsets of TFs important for predicting cell-cycle expression. Finally, analysis of important TF-TF interaction features suggests that the GRN regulating cell cycle expression is highly interconnected and clustered around four groups of genes, two of which represent known cell-cycle regulatory complexes, while the other two contain TFs that are not known cell-cycle regulators (Ste12-Tex1 and Rap1-Hap1-Msn4), but are nonetheless important to regulating the timing of expression. Thus, not only do our models accurately reflect what is known about the regulation of the S. cerevisiae cell cycle, they can be used to discover regulatory factors which play a role in controlling expression during the cell cycle as well as other contexts with discrete temporal patterns of expression.

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Total Phenols, Identification of Active Compounds and Anticancer Activity of Salvia judaica Boiss against the breast Cancer Cell MDA-231

Journal of Bioscience & Biomedical Engineering ◽

10.47485/2693-2504.1052 ◽

2021 ◽

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Essential Oil ◽

Anticancer Activity ◽

Breast Cancer Cell ◽

Annexin V ◽

Ethanol Extract ◽

G1 Phase ◽

Apoptotic Cells ◽

Total Phenols

Salvia judaica is an annual herb from genus Salvia L.; the largest genera of Lamiaceae. It’s a medicinal plant prominent in pharmaceutical applications in many countries around the world. This study aimed to explore bioactive compounds likely to be responsible for the plant anticancer activity, and evaluate anticancer effects, after determining the total content of phenols in the ethanol extract and essential oil in this species. Ethanol extract (EE) and essential oil (EO) were prepared from dried aerial parts (leaves and the flower). GC-MS analysis of EO showed the presence of/43/ effective compounds in varying proportions, the major compounds were sesquiterpenes like delta-cadinene, alpha-Gurjunene, beta-humulene, and alpha-caryophyllene. This is the first study revealed that S.judaica is so rich in phenols which proceeded S.officinalis, noting the superiority of the EE over the EO samples in the total phenols. Anticancer properties of EE and EO of S. judaica against MDA-231 breast cancer cell line were studied -for the first time - by cell cycle analysis and Annexin V/PI apoptosis assay using Flow cytometry technique. Cells were treated with EE (0.001, 0.01, 0.02, 0.1mg/ml) and EO (0.005, 0.01, 0.02, 0.03, 0.04 mg/ml) at various concentrations for48 h. The results revealed that both EE and EO induced cell cycle arrest at G1-phase. Cells treated with EE and EO for 48h showed increasing the percentage of cells in G1-phase and decreasing the percentage of cells in S-phase with increasing concentration compared with untreated cells (control). Annexin V-FITC/PI assay confirmed that EO and EE were able to induce apoptosis. Cells treated with EOat (0.04 mg/ml) for 48h resulted in apoptotic cells at 96.68%, and necrotic cells at 0.12%, compared with untreated cells. On the other hand, Cells treated with EE at (0.1 mg/ml) for 48h resulted in apoptotic cells at 94.43%, and necrotic cells at 0.47%, compared with control. Results revealed that EO is better than EE as anticancer; treatment with EO resulted in more apoptotic cells and less necrotic cells, and there were significant differences between them. This confirmed that EO contains specific anticancer compounds as showed by GC-MS analysis. However, more studies should be performed to explore antioxidants present in S.judaica and determine the underlying mechanism of their anti-breast cancer properties.

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Induction of apoptosis by Kola nut extract as a recent and promising treatment strategy for Leukemia

Bionatura ◽

10.21931/rb/2021.06.02.10 ◽

2021 ◽

Vol 6 (2) ◽

pp. 1725-1732

Author(s):

Hamdah Alsaeedi ◽

Rowaid Qahwaji ◽

Talal Qadah

Keyword(s):

Cell Cycle ◽

Cell Line ◽

Leukemia Cell ◽

K562 Cells ◽

Myelogenous Leukemia ◽

Time Dependent ◽

Leukemia Cell Line ◽

Apoptotic Cells ◽

Dependent Manner ◽

Anti Cancer

Kola nut extracts have recently been reported to contain chemopreventive compounds providing several pharmacological benefits. This study investigated Kola nut extracts' anti-cancer activity on human immortalized myelogenous leukemia cell line K562 through apoptosis and cell cycle arrest. Fresh Kola nuts were prepared as powder and dissolved in DMSO. Different concentrations (50, 100, 150, 200, and 250 μg/ml) of working solutions were prepared. The K562 cells were treated with the different concentrations of Kola nut extract or vehicle control (10% DMSO) followed by incubation at 37°C for 24, 48, and 72 hours, respectively. Treatment activity was investigated in K562 cells; by Resazurin, and FITC/Propidium Iodide and 7-AAD stained cells to evaluate apoptotic cells and the cell cycle's progression. Inhibition of leukemia cell proliferation was observed. The extract effectively induced cell death, early and late apoptosis by approximately 30% after 24 and 48 hours incubation, and an increase in the rate of dead cells by 50% was observed after 72 hours of incubation. Also, cell growth reduction was seen at high dose concentrations (150 and 200 µg/ml), as evident by cell count once treated with Kola nut extract. The total number of apoptotic cells increased from 5.8% of the control group to 27.4% at 250 µg/ml concentration. Moreover, Kola nut extracts' effects on K562 cells increased gradually in a dose and time-dependent manner. It was observed that Kola nut extracts could arrest the cell cycle in the G2/M phase as an increase in the number of cells by 29.8% and 14.6 % were observed from 9.8% and 5.2% after 24 and 48 hours of incubation, respectively. This increase was detected in a dose and time-dependent manner. Kola nut extracts can be used as a novel anti-cancer agent in Leukemia treatment as it has shown significant therapeutic potential and therefore provides new insights in understanding the mechanisms of its action. Keywords: Kola nut extracts, Leukemia, K562 cell line, Apoptosis, Cancer.

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