scholarly journals Deciphering colchicine like actions of clerodin in terms of microtubule destabilization based mitotic abnormalities, G2/M-phase arrest, and plant polyploidy

2020 ◽  
Author(s):  
Sujit Roy ◽  
Lalit Mohan Kundu ◽  
Gobinda Chandra Roy ◽  
Manabendu Barman ◽  
Tista Chakraborty ◽  
...  
Keyword(s):  
Molecular Docking ◽  
Human Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Root Apical Meristem ◽  
Root Tip ◽  
Common Mechanism ◽  
Microscopy Imaging ◽  
Flow Cytometric ◽  
Mitotic Abnormalities ◽  
Mcf 7

AbstractClerodin (C24H34O7), a clerodane diterpenoid, is a bitter principle of Clerodendrum viscosum. The present study aimed to decipher colchicine-like actions of clerodin in terms of microtubule destabilization based mitotic abnormalities, G2-M arrest, and plant polyploidy. Purified clerodin showed increased metaphase frequency in Human Peripheral Blood Lymphocytes (HPBLs), Human Embryonic Kidney cells (HEK-293), and Allium cepa root apical meristem cells. Both squashed slide of the onion root tip and flow cytometric analysis of radish protoplast revealed a significantly increased frequency of polyploid cells. Flow cytometric analysis showed an increase in frequencies of G2-M in MCF-7 cells from 6.10 to 16.25% after clerodin (200μg/mL) treatment for 24 h. Confocal microscopy imaging of tubulin in clerodin-treated MCF-7 cells revealed microtubule destabilization. Molecular docking and LIGPLOT analysis indicate that clerodin interact in the colchicine binding site, including, single hydrogen bond with Asn 101 of α-tubulin. In summary, our experimental data revealed that clerodin has metaphase arresting, microtubule destabilization, and polyploidy inducing ability similar to colchicine. Molecular docking analysis revealed for the first time that clerodin and colchicine interact at the common site of tubulin residue indicating a common mechanism of action. The results also indicate similar cytotoxic potentialities of both clerodin and colchicine even though they belong to different chemical groups. Thus, clerodin may be used in place of colchicine as a plant polyploidy inducing agent in plant breeding programs in Agriculture.

Cell synchronization and isolation of metaphase chromosomes from maize (Zea mays L.) root tips for flow cytometric analysis and sorting

Genome ◽  
1996 ◽  
Vol 39 (4) ◽  
pp. 697-703 ◽  
Author(s):  
Jai-Heon Lee ◽  
K. Arumuganathan ◽  
S. M. Kaeppler ◽  
C. M. Papa ◽  
H. F. Kaeppler
Keyword(s):  
Cell Cycle ◽  
Flow Cytometric Analysis ◽  
Maize Root ◽  
Root Tip ◽  
Root Tips ◽  
Chromosome Sorting ◽  
Synthesis Inhibitor ◽  
Metaphase Chromosomes ◽  
Flow Karyotype ◽  
Flow Cytometric

Accumulation of cells containing metaphase chromosomes is an important step in cytological analyses and chromosome sorting procedures. The goal of this research was to optimize treatment parameters to synchronize the cell cycle of maize root tip meristem cells. Levels of hydroxyurea, a DNA synthesis inhibitor, were assessed for their utility in accumulating cells at the G1 phase of the cell cycle. Trifluralin, amiprophos-methyl, and colchicine were used to accumulate cells containing metaphase chromosomes upon release from hydroxyurea inhibition. Optimal mitotic indices were achieved by treating seedlings with 5 mM hydroxyurea for 18 h, incubating for 1 h without chemical treatment to release the hydroxyurea block, and then treating emerging roots with 1 μM trifluralin for 4 h. The mitotic index of synchronized maize root tips was over 70%. Uniformity of synchronization depended upon selection of seeds with emerging radicles that were similar in length at the time of treatment. Suspensions of intact chromosomes were prepared by a simple slicing procedure. The chromosome preparations were found to be suitable for flow cytometric characterization and sorting. Chromosome peaks of the observed flow karyotype resembled the predicted flow karyotype calculated on the basis of maize chromosome size. Key words : flow karyotype, hydroxyurea, plant chromosome sorting, trifluralin.

scholarly journals Polyploidy in Stokes Aster (Stokesia laevis)

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1101A-1101
Author(s):  
Jessica Gaus ◽  
Dennis Werner ◽  
Shyamalrau Tallury
Keyword(s):  
Segregation Analysis ◽  
Karyotype Analysis ◽  
Flow Cytometric Analysis ◽  
Root Tip ◽  
Hybrid Progeny ◽  
Chromosome Counts ◽  
Segregation Behavior ◽  
Flow Cytometric ◽  
Triploid Block ◽  
Aberrant Segregation

Segregation analysis of two different F2 families of stokes aster created by hybridizing two blue-flowered cultivars [`Peaches Pick' (PE) and `Omega Skyrocket' (OSR)] with the yellow-flowered cultivar `Mary Gregory' (MG) gave disparate results. The F2 progeny of PE × MG segregated in the expected 3:1 (blue:yellow) ratio. In contrast, all 782 progeny from the MG × OSR F2 family were blue-flowered. Flow cytometric analysis of the parents and F1 hybrids was conducted to determine if ploidy differences existed among the parents, as such differences could account for aberrant segregation behavior in the MG × OSR F2 family. Peak ratios suggested that MG and PE were diploid, OSR was tetraploid, and F1 hybrids of MG × OSR were triploid. Chromosome counts from root tip squashes confirmed that MG and PE were diploid (2n= 2x= 14), OSR was tetraploid (2n= 4x= 28), and F1 hybrid progeny of MG × OSR were triploid (2n= 3x= 21). Karyotype analysis also confirmed these results. We propose that the lack of recovery of yellow-flowered progeny in the MG × OSR F2 family is due to differences in parental chromosome number. These results document the first report of polyploidy in stokes aster, and suggest the absence of a triploid block in this species.

Interferon gamma delays apoptosis of mature erythroid progenitor cells in the absence of erythropoietin

Blood ◽  
2000 ◽  
Vol 95 (12) ◽  
pp. 3742-3749 ◽  
Author(s):  
Ilseung Choi ◽  
Koichiro Muta ◽  
Amittha Wickrema ◽  
Sanford B. Krantz ◽  
Junji Nishimura ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Interferon Gamma ◽  
Human Peripheral Blood ◽  
Flow Cytometric Analysis ◽  
Annexin V ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cells ◽  
Flow Cytometric ◽  
Ifn Γ ◽  
Protease Assay

Based on the hypothesis that interferon gamma (IFN-γ) may have stimulating effects on survival of hematopoietic progenitor cells, we examined the effect of IFN-γ on apoptosis of mature erythroid colony-forming cells (ECFCs) derived from human peripheral blood obtained from normal, healthy volunteers. When the cells were cultured in the presence of IFN-γ, even without erythropoietin (EPO), the viability of the cells was maintained for at least 36 hours. When apoptosis of ECFCs was assessed by flow cytometric analysis', using annexin V, IFN-γ reduced the extent of apoptosis of the cells, as well as EPO. DNA fragmentation of ECFCs was also reduced by IFN-γ. In cells cultured with IFN-γ alone, expression of Bcl-x was detected but the level of expression decreased gradually during incubation for 36 hours, and the expression level was lower than incubation with EPO. Fas expression and activation of downstream caspases were assessed by flow cytometric analysis or fluorometric protease assay. IFN-γ induced Fas expression of the cells without the activation of caspase8 or caspase3 during 16 hours of incubation, while deprivation of EPO induced expression of Fas and the activation of both caspase8 and caspase3. We propose that IFN-γ produces a stimulating signal for the survival of mature erythroid progenitor cells by reducing apoptosis through a mechanism other than modulating Fas and one related to the expression of Bcl-x.

scholarly journals Synthesis, Anti-Tumor Activity and Apoptosis-Inducing Effect of Novel Dimeric Keggin-Type Phosphotungstate

2021 ◽  
Vol 11 ◽  
Author(s):  
Yingxue Xue ◽  
Yifei Yin ◽  
He Li ◽  
Mingyu Chi ◽  
Jiaxin Guo ◽  
...  
Keyword(s):  
Flow Cytometric Analysis ◽  
A549 Cells ◽  
Antitumor Action ◽  
Dependent Manner ◽  
Keggin Type ◽  
Flow Cytometric ◽  
Ft Ir ◽  
Octadecyltrimethylammonium Bromide ◽  
Mcf 7

A dimeric Keggin-type phosphotungstate (ODA)10[(PW11FeO39)2O]·9H2O (abbreviated as ODA10[(PW11Fe)2], ODA = octadecyltrimethylammonium bromide) was synthesized and investigated comprehensively its antitumor activity on MCF-7 and A549 cells. The dimeric structure and amorphous morphology were characterized by FT-IR, UV-vis-DRS, SEM and XRD. The in vitro MTT assay of ODA10[(PW11Fe)2] showed anticancer activity on MCF-7 and A549 cells in a dose- and time-dependent manner, and the IC50 values for MCF-7 and A549 cells at 48 h were 5.83 μg/ml and 3.23 μg/ml, respectively. The images of the ODA10[(PW11Fe)2]-treated cells observed by inverted biological microscope exhibited the characteristic morphology of apoptosis. Flow cytometric analysis showed cell apoptosis and cycle arrested at S phase induced by ODA10[(PW11Fe)2]. The above results illuminated the main mechanism of the antitumor action of ODA10[(PW11Fe)2] on MCF-7 and A549 cells, indicating that this dimeric phosphotungstate is a promising anticancer drug.

The Role of Beclin1 Gene in Autophagy and Apoptosis Induced by Ionizing Radiation in MCF - 7 Cells

2013 ◽  
Vol 790 ◽  
pp. 587-589
Author(s):  
Ya Li Qi ◽  
Yan Jun Liu ◽  
Da Li Zhao
Keyword(s):  
Ionizing Radiation ◽  
Protein Expression ◽  
Flow Cytometric Analysis ◽  
Beclin 1 ◽  
Over Expression ◽  
Western Blot Method ◽  
Flow Cytometric ◽  
Dose Effects ◽  
Mcf 7

Objective To discuss the role of Beclin1 gene in autophagy and apoptosis induced by ionizing radiation in MCF - 7 cells. Methods MTT assay was used to detect the influence of cells proliferation when beclin-1 gene was over expression and interference. Flow cytometric analysis detected the change of MCF - 7 cells apoptosis after irradiating by X-ray. Western blot method detected total protein changes of beclin - 1. Results Beclin-1 gene was interferred partly, the amount of protein expression of MCF-7-beclin1Ri cells reached to the lowest at 4 h, rose at 8 h, got to the most at 16 h and decreased at 32 h, but still held at higher level;Beclin-1 gene over-expression, the amount of protein expression increased gradually with the extension of time, and got to peak at 32 h. Conclusions Ionizing radiation can stimulate beclin-1 and exist relations of dose - effects in MCF-7 cells.

Sign in / Sign up

Export Citation Format

Share Document