scholarly journals CYTOTOXICITY EFFECT OF IONIC LIQUID-GRAVIOLA FRUIT (ANNONA MURICATA) EXTRACT TO HUMAN COLON CANCER (HT29) CELL LINES

2021 ◽  
Vol 22 (2) ◽  
pp. 50-66
Author(s):  
Djabir Daddiouaissa ◽  
Azura Amid ◽  
NASSERELDEEN AHMED KABBASHI ◽  
AHMED ADAM MOHAMMED ELNOUR ◽  
MOHAMAD ADIKA KHAIRY BIN MOHD SHAIFUDIN EPANDY
Keyword(s):  
Cell Cycle ◽  
Ionic Liquid ◽  
Cell Lines ◽  
Colon Adenocarcinoma ◽  
Ht29 Cell ◽  
G1 Phase ◽  
Cycle Phase ◽  
Human Colon Cancer ◽  
Ht29 Cells ◽  
Ic50 Value

The present study aimed to investigate the anti-proliferative effect of the ionic liquid-Graviola fruit (IL-GFE) extract on colon adenocarcinoma (HT29) cell lines and their kinetics behaviour to assess the Graviola fruit potential as a therapeutic alternative in cancer treatment. The phytoconstituents content of IL-GFE was identified using GC-TOFMS apparatus and measured its cytotoxicity on HT29 by tetrazolium bromide. Then the cytokinetic behaviour of the treated HT29 cells with IL-GFE was illustrated using the cells' growth curve. Besides, the cell cycle phase perturbation for the treated HT29 was applied using a flow cytometry technique. Qualitative identification of phytoconstituents of IL-GFE showed that Graviola fruit contains acetogenins, alkaloids, flavonoids, tannins and saponins compounds. IL-GF extract displayed a cytotoxicity effect on HT29 cells with the IC50 value of 10.56 µg/mL, while Taxol showed an IC50 value of 1.22 µg/mL. IL-GFE also decreased the cell generation number from 3.93 to 2.96 generations compared to Taxol-treated cells 2.01 generations. The microscope observation of the HT29 cells treated with the crude IL-GFE displayed loss of density and cell detachment. The extract's growth inhibition was related to the cell cycle arrest at the G0/G1 phase. IL-GFE inhibited colon adenocarcinoma HT29 cells' proliferation and affected their kinetic behaviour by lowering cell viability, inducing apoptosis, and arresting the cell cycle at the G0/G1 phase. ABSTRAK: Kajian ini bertujuan untuk mengkaji kesan anti-proliferatif ekstrak buah-ion Graviola (IL-GFE) pada garis sel adenokarsinoma kolon (HT29) dan tingkah laku kinetik mereka untuk menilai potensi buah Graviola sebagai alternatif terapi untuk barah rawatan. Kandungan fitokonstituen IL-GFE dikenal pasti menggunakan alat GC-TOFMS dan mengukur sitotoksisitasnya pada HT29 oleh tetrazolium bromida. Kemudian tingkah laku sitokinetik sel HT29 yang dirawat dengan IL-GFE digambarkan menggunakan keluk pertumbuhan sel. Selain itu, gangguan fasa kitaran sel untuk HT29 yang dirawat diaplikasikan menggunakan teknik sitometri aliran. Pengenalpastian kualitatif fitokonstituen IL-GFE menunjukkan bahawa buah Graviola mengandungi asetogenin, alkaloid, flavonoid, tanin dan sebatian saponin. Ekstrak IL-GF memperlihatkan kesan sitotoksisiti pada sel HT29 dengan nilai IC50 10.56 µg/mL, sementara Taxol menunjukkan nilai IC50 1.22 µg/mL. IL-GFE juga menurunkan jumlah penjanaan sel dari 3.93 hingga 2.96 generasi berbanding sel yang dirawat Taxol 2.01 generasi. Pemerhatian mikroskop sel HT29 yang dirawat dengan IL-GFE kasar menunjukkan kehilangan ketumpatan dan detasmen sel. Perencatan pertumbuhan ekstrak berkaitan dengan penangkapan kitaran sel pada fasa G0/G1. IL-GFE menghalang percambahan sel HT29 adenokarsinoma kolon dan mempengaruhi tingkah laku kinetik mereka dengan menurunkan daya maju sel, mendorong apoptosis, dan menghentikan kitaran sel pada fasa G0/G1.

scholarly journals Evaluation of metabolomics behavior of human colon cancer HT29 cell lines treated with ionic liquid graviola fruit pulp extract

2021 ◽  
Vol 270 ◽  
pp. 113813
Author(s):  
Djabir Daddiouaissa ◽  
Azura Amid ◽  
Muhamad Shirwan Abdullah Sani ◽  
Ahmed A.M. Elnour
Keyword(s):  
Colon Cancer ◽  
Ionic Liquid ◽  
Cell Lines ◽  
Human Colon ◽  
Ht29 Cell ◽  
Fruit Pulp ◽  
Human Colon Cancer

scholarly journals Effect of cell cycle position on dexamethasone binding by mouse and human lymphoid cell lines: correlation between an increase in dexamethasone binding during S phase and dexamethasone sensitivity

Blood ◽  
1984 ◽  
Vol 63 (1) ◽  
pp. 105-113
Author(s):  
CW Distelhorst ◽  
BM Benutto ◽  
RA Bergamini
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Lymphoid Cell ◽  
S Phase ◽  
Lymphoid Cells ◽  
Cell Protein ◽  
G1 Phase ◽  
Cycle Phase ◽  
Human Lymphoid Cell ◽  
Lymphoid Cell Lines

We determined the effect of cell cycle position on the amount of dexamethasone that was specifically bound by mouse and human lymphoid cell lines. Cell lines that were either sensitive or resistant to growth inhibition by dexamethasone were compared. Exponentially growing cells were separated by centrifugal elutriation into fractions that corresponded to different positions in the cell cycle. The cell cycle phase distribution of each fraction was estimated by flow cytometry and autoradiography. The amount of dexamethasone bound per cell in each fraction was measured by a whole cell binding assay. In three dexamethasone-sensitive cell lines (two mouse and one human), we found that the amount of dexamethasone bound per cell increased 2–4-fold between G1 phase and S phase, and then decreased during G2/M phase. Results were the same when the amount of dexamethasone bound per milligram of cell protein was measured. Binding affinity was the same during G1 phase and S phase, but the proportion of bound dexamethasone that translocated to the nucleus was greater during S phase. In contrast, we found that the amount of dexamethasone bound per cell by three dexamethasone-resistant cell lines (two mouse and one human) did not increase during S phase. Our results indicate that cell cycle changes in dexamethasone binding are not simply related to changes in cell protein or cell volume during the cell cycle. An increase in dexamethasone binding during S phase may be required for dexamethasone to inhibit cell growth, and a failure of dexamethasone binding to increase during S phase might represent a new mechanism of dexamethasone resistance in lymphoid cells.

scholarly journals Effect of cell cycle position on dexamethasone binding by mouse and human lymphoid cell lines: correlation between an increase in dexamethasone binding during S phase and dexamethasone sensitivity

Blood ◽  
1984 ◽  
Vol 63 (1) ◽  
pp. 105-113 ◽  
Author(s):  
CW Distelhorst ◽  
BM Benutto ◽  
RA Bergamini
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Lymphoid Cell ◽  
S Phase ◽  
Lymphoid Cells ◽  
Cell Protein ◽  
G1 Phase ◽  
Cycle Phase ◽  
Human Lymphoid Cell ◽  
Lymphoid Cell Lines

Abstract We determined the effect of cell cycle position on the amount of dexamethasone that was specifically bound by mouse and human lymphoid cell lines. Cell lines that were either sensitive or resistant to growth inhibition by dexamethasone were compared. Exponentially growing cells were separated by centrifugal elutriation into fractions that corresponded to different positions in the cell cycle. The cell cycle phase distribution of each fraction was estimated by flow cytometry and autoradiography. The amount of dexamethasone bound per cell in each fraction was measured by a whole cell binding assay. In three dexamethasone-sensitive cell lines (two mouse and one human), we found that the amount of dexamethasone bound per cell increased 2–4-fold between G1 phase and S phase, and then decreased during G2/M phase. Results were the same when the amount of dexamethasone bound per milligram of cell protein was measured. Binding affinity was the same during G1 phase and S phase, but the proportion of bound dexamethasone that translocated to the nucleus was greater during S phase. In contrast, we found that the amount of dexamethasone bound per cell by three dexamethasone-resistant cell lines (two mouse and one human) did not increase during S phase. Our results indicate that cell cycle changes in dexamethasone binding are not simply related to changes in cell protein or cell volume during the cell cycle. An increase in dexamethasone binding during S phase may be required for dexamethasone to inhibit cell growth, and a failure of dexamethasone binding to increase during S phase might represent a new mechanism of dexamethasone resistance in lymphoid cells.

scholarly journals Diclofenac N-Derivatives as Therapeutic Agents with Anti-Inflammatory and Anti-Cancer Effect

2021 ◽  
Vol 22 (10) ◽  
pp. 5067
Author(s):  
Alberto Galisteo ◽  
Fatin Jannus ◽  
Amalia García-García ◽  
Houssam Aheget ◽  
Sara Rojas ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Ht29 Cell ◽  
Cycle Phase ◽  
No Production ◽  
Hep G2 ◽  
Anti Cancer ◽  
Anti Inflammatory

A series of diclofenac N-derivatives (2, 4, 6, 8c, 9c, 10a-c) were synthesized in order to test their anti-cancer and anti-inflammatory effects. The anticarcinogen activity has been assayed against three cancer cell lines: HT29, human colon cancer cells; Hep-G2, human hepatic cells; and B16-F10, murine melanoma cells. First, we determined the cytotoxicity of the different compounds, finding that the most effective compound was compound 8c against all cell lines and both compounds 4 and 6 in human Hep-G2 and HT29 cell lines. Compounds 4 and 8c were selected for the percentage of apoptosis determination, cell cycle distribution, and mitochondrial membrane potential measure because these products presented the lowest IC50 values in two of the three cancer cell lines assayed (B16-F10 and HepG2), and were two of the three products with lowest IC50 in HT29 cell line. Moreover, the percentages of apoptosis induction were determined for compounds 4 and 8c, showing that the highest values were between 30 to 60%. Next, the effects of these two compounds were observed on the cellular cycle, resulting in an increase in the cell population in G2/M cell cycle phase after treatment with product 8c, whereas compound 4 increased the cells in phase G0/G1, by possible differentiation process induction. Finally, to determine the possible apoptosis mechanism triggered by these compounds, mitochondrial potential was evaluated, indicating the possible activation of extrinsic apoptotic mechanism. On the other hand, we studied the anti-inflammatory effects of these diclofenac (DCF) derivatives on lipopolysaccharide (LPS) activated RAW 264.7 macrophages-monocytes murine cells by inhibition of nitric oxide (NO) production. As a first step, we determined the cytotoxicity of the synthesized compounds, as well as DCF, against these cells. Then, sub-cytotoxic concentrations were used to determine NO release at different incubation times. The greatest anti-inflammatory effect was observed for products 2, 4, 8c, 10a, 10b, and 9c at 20 µg·mL−1 concentration after 48 h of treatment, with inhibition of produced NO between 60 to 75%, and a concentration that reduces to the 50% the production of NO (IC50 NO) between 2.5 to 25 times lower than that of DCF. In this work, we synthesized and determined for the first time the anti-cancer and anti-inflammatory potential of eight diclofenac N-derivatives. In agreement with the recent evidences suggesting that inflammation may contribute to all states of tumorigenesis, the development of these new derivatives capable of inducing apoptosis and anti-inflammatory effects at very low concentrations represent new effective therapeutic strategies against these diseases.

scholarly journals Resistance of Hypoxic Cells to Ionizing Radiation Is Mediated in Part via Hypoxia-Induced Quiescence

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 610
Author(s):  
Apostolos Menegakis ◽  
Rob Klompmaker ◽  
Claire Vennin ◽  
Aina Arbusà ◽  
Maartje Damen ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Cancer Cell ◽  
Radiation Treatment ◽  
Large Fraction ◽  
Radiation Sensitivity ◽  
Cancer Cell Lines ◽  
Cycle Phase ◽  
Multicellular Spheroids ◽  
Phase Duration

Double strand breaks (DSBs) are highly toxic to a cell, a property that is exploited in radiation therapy. A critical component for the damage induction is cellular oxygen, making hypoxic tumor areas refractory to the efficacy of radiation treatment. During a fractionated radiation regimen, these hypoxic areas can be re-oxygenated. Nonetheless, hypoxia still constitutes a negative prognostic factor for the patient’s outcome. We hypothesized that this might be attributed to specific hypoxia-induced cellular traits that are maintained upon reoxygenation. Here, we show that reoxygenation of hypoxic non-transformed RPE-1 cells fully restored induction of DSBs but the cells remain radioresistant as a consequence of hypoxia-induced quiescence. With the use of the cell cycle indicators (FUCCI), cell cycle-specific radiation sensitivity, the cell cycle phase duration with live cell imaging, and single cell tracing were assessed. We observed that RPE-1 cells experience a longer G1 phase under hypoxia and retain a large fraction of cells that are non-cycling. Expression of HPV oncoprotein E7 prevents hypoxia-induced quiescence and abolishes the radioprotective effect. In line with this, HPV-negative cancer cell lines retain radioresistance, while HPV-positive cancer cell lines are radiosensitized upon reoxygenation. Quiescence induction in hypoxia and its HPV-driven prevention was observed in 3D multicellular spheroids. Collectively, we identify a new hypoxia-dependent radioprotective phenotype due to hypoxia-induced quiescence that accounts for a global decrease in radiosensitivity that can be retained upon reoxygenation and is absent in cells expressing oncoprotein E7.

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

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2569 ◽  
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.

Cyclin D1 Maintains Mantle Cell Lymphoma Through CDK4-Independent Regulation Of DNA Replicative Checkpoints

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2512-2512
Author(s):  
Suchismita Mohanty ◽  
Atish Mohanty ◽  
Natalie Sandoval ◽  
Victoria Bedell ◽  
Joyce Murata-Collins ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Survival ◽  
Cell Lines ◽  
Cell Lymphoma ◽  
Replication Stress ◽  
G1 Phase ◽  
Origin Firing ◽  
Mantle Cell ◽  
Cdk4 Activity

Abstract Mantle cell lymphoma (MCL) is rarely curable and therapy resistance often leaves few viable treatment options for patients. Previous studies have identified the importance of cyclin D1 (CCND1) translocation and overexpression in MCL pathogenesis, which leads to increased cyclin-dependent kinase 4 (CDK4) activity and accelerated cell cycle progression. However, targeting this abnormal cell cycle control, mainly through CDK4 inhibition causes only G1-phase growth arrest without significant cell death (Marzec et al. 2006). In contrast, prolonged inhibition of CCND1 with RNA interference induces apoptosis in MCL cell lines (Weinstein et al. 2012), suggesting an essential function of CCND1 independent of CDK4 activity. The mechanism of this non-catalytic role of CCND1 in maintaining MCL cell survival is largely unknown. To clarify the cell cycle role of CCND1 in addition to its CDK4-dependent function, we compared the effects of CCND1 and CDK4 silencing on MCL cell survival. MCL cell lines co-expressing GFP and doxycycline-inducible shRNA targeting CCND1 or CDK4 were generated. Cells with similar GFP expression levels were FACS sorted to normalize for shRNA expression. Both CCND1 and CDK4 silencing resulted in G1-phase arrest, but only CCND1-silenced cells demonstrated a marked increase in apoptosis. Investigation of the potential cause of apoptosis revealed significant accumulation of DNA double-strand breaks following CCND1 ablation, as measured by nuclear gamma-H2AX focus formation. Interestingly, CCND1-silenced cells exhibited a significant increase in 53BP1+ nuclear bodies in G1-phase, reminiscent of 53BP1 foci observed by Lukas and colleagues in cells undergoing aphidicolin-induced replication stress (Lukas et al. 2011). Analysis of replication fork movement in CCND1-depleted cells showed substantially reduced fork speed and increased frequency of origin firing, both of which are indicative of replication stress. In contrast, knockdown of CDK4 did not result in slower forks or increase in the frequency of origin firing. Genomic instability associated with replication stress was also apparent in CCND1-silenced cells, including increased micronucleus formation and recurrent chromatid gaps or breaks detected by cytokinesis-block assay and karyotyping, respectively. Analysis of DNA replicative and damage checkpoints revealed that both ATR-CHEK1 and ATM-CHEK2 pathways were activated by phosphorylation following CCND1 silencing in MCL cell lines, a xenograft animal model, and primary tumor samples, but not in non-MCL tumors. Interestingly, this activation (with the exception of ATM phosphorylation) was unsustainable over time and did not cause down-regulation of the downstream targets CDC25 and CDK1/2 but, instead, we observed an increase in CDC25A/B protein levels and CDK1/2 activity, indicating defective cell cycle checkpoints. Exposing CCND1-silenced cells to replication stress-inducing or DNA-damaging agents such hydroxyurea, aphidicolin, etoposide or ionizing radiation further amplified the checkpoint defects seen in unperturbed cells. We did not observe any significant difference in this checkpoint signaling in control and CDK4 knockdown cells under these conditions. Furthermore, CCND1-deficient cells were more sensitive to pharmacological inhibition of ATR and CHEK1 but not ATM, confirming a constitutive role of CCND1 in the ATR-CHEK1 pathway. In conclusion, these studies revealed an unexpected CDK4-independent role of CCND1 in maintaining DNA replicative checkpoints to prevent replication stress and genome instability in MCL cells. As most cancer treatments rely on agents that create DNA replication stress, targeting this function of CCND1 could provide a rational approach to overcome resistance to conventional therapies in MCL. Disclosures: No relevant conflicts of interest to declare.

scholarly journals A mathematical model of subpopulation kinetics for the deconvolution of leukaemia heterogeneity

2015 ◽  
Vol 12 (108) ◽  
pp. 20150276 ◽  
Author(s):  
María Fuentes-Garí ◽  
Ruth Misener ◽  
David García-Munzer ◽  
Eirini Velliou ◽  
Michael C. Georgiadis ◽  
...  
Keyword(s):  
Mathematical Model ◽  
Cell Cycle ◽  
Cell Lines ◽  
Phase Distribution ◽  
Initial Conditions ◽  
Cellular Heterogeneity ◽  
Cycle Phase ◽  
Biologically Relevant ◽  
Future Outcomes ◽  
Unique Cell

Acute myeloid leukaemia is characterized by marked inter- and intra-patient heterogeneity, the identification of which is critical for the design of personalized treatments. Heterogeneity of leukaemic cells is determined by mutations which ultimately affect the cell cycle. We have developed and validated a biologically relevant, mathematical model of the cell cycle based on unique cell-cycle signatures, defined by duration of cell-cycle phases and cyclin profiles as determined by flow cytometry, for three leukaemia cell lines. The model was discretized for the different phases in their respective progress variables (cyclins and DNA), resulting in a set of time-dependent ordinary differential equations. Cell-cycle phase distribution and cyclin concentration profiles were validated against population chase experiments. Heterogeneity was simulated in culture by combining the three cell lines in a blinded experimental set-up. Based on individual kinetics, the model was capable of identifying and quantifying cellular heterogeneity. When supplying the initial conditions only, the model predicted future cell population dynamics and estimated the previous heterogeneous composition of cells. Identification of heterogeneous leukaemia clones at diagnosis and post-treatment using such a mathematical platform has the potential to predict multiple future outcomes in response to induction and consolidation chemotherapy as well as relapse kinetics.

Induction of apoptosis in K562 cells by jolkinolide B

2006 ◽  
Vol 84 (10) ◽  
pp. 959-965 ◽  
Author(s):  
Huiying Luo ◽  
Aiqin Wang
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
K562 Cells ◽  
G1 Phase ◽  
Fragmentation Pattern ◽  
Nuclear Condensation ◽  
Transmission Electron ◽  
Agarose Electrophoresis ◽  
Euphorbia Fischeriana ◽  
Hepatoma Hepg2

Jolkinolide B, a bioactive diterpene isolated from the roots of Euphorbia fischeriana Steud, has various biological and pharmacological properties. In this study, the cytotoxicity of highly purified jolkinolide B was tested in human chronic myeloid leukemia (K562) and 2 other cell lines (human esophageal carcinoma Eca-109 and human hepatoma HepG2). The results indicate a significant decrease in the proliferation of all the 3 cell lines when treated with jolkinolide B for 24 h; the IC50 value of cytotoxicity was 12.1 μg/mL (for K562 cells), >50.0 μg/mL (for HepG2 cells), and 23.7 μg/mL (for Eca-109 cells). Further study of K562 cells involving fluorescence and transmission electron microscopy revealed characteristic apoptotic features, such as cell shrinkage, membrane blebbing, loss of microvilli, and nuclear condensation. Agarose electrophoresis of genomic DNA showed a typical fragmentation pattern for apoptotic cells. A kinetic cell-cycle analysis demonstrated that the cell cycle was arrested in the G1 phase. All these results suggest that the anti-proliferation effect of jolkinolide B on K562 cells is achieved by arresting the cell cycle in the G1 phase and subsequently inducing apoptosis.

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