scholarly journals Improved Split-GFP Systems for Visualizing Organelle Contact Sites in Yeast and Human Cells

2020 ◽  
Vol 8 ◽  
Author(s):  
Shinya Tashiro ◽  
Yuriko Kakimoto ◽  
Manatsu Shinmyo ◽  
Shintaro Fujimoto ◽  
Yasushi Tamura
Keyword(s):  
Hela Cell ◽  
Cell Line ◽  
Mammalian Cells ◽  
Cultured Cells ◽  
Early Stage ◽  
Human Cells ◽  
Yeast Cells ◽  
Hela Cell Line ◽  
Contact Sites ◽  
Low Levels

Inter-organelle contact sites have attracted a lot of attention as functionally specialized regions that mediate the exchange of metabolites, including lipids and ions, between distinct organelles. However, studies on inter-organelle contact sites are at an early stage and it remains enigmatic what directly mediates the organelle-organelle interactions and how the number and degree of the contacts are regulated. As a first step to answer these questions, we previously developed split-GFP probes that could visualize and quantify multiple inter-organelle contact sites in the yeast and human cultured cells. However, the split-GFP probes possessed a disadvantage of inducing artificial connections between two different organelle membranes, especially when overexpressed. In the present study, we developed a way to express the split-GFP probes whose expressions remained at low levels, with minimal variations between different yeast cells. Besides, we constructed a HeLa cell line in which the expression of the split-GFP probes could be induced by the addition of doxycycline to minimize the artificial effects. The improved split-GFP systems may be faithful tools to quantify organelle contact sites and screen new factors involved in organelle-organelle tethering in yeast and mammalian cells.

Glyco-gold nanoparticle shapes enhance carbohydrate–protein interactions in mammalian cells

Nanoscale ◽  
2016 ◽  
Vol 8 (25) ◽  
pp. 12729-12735 ◽  
Author(s):  
Sivakoti Sangabathuni ◽  
Raghavendra Vasudeva Murthy ◽  
Preeti Madhukar Chaudhary ◽  
Manalee Surve ◽  
Anirban Banerjee ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Hela Cell ◽  
Cell Line ◽  
Gold Nanoparticle ◽  
Protein Interactions ◽  
Mammalian Cells ◽  
Hela Cell Line ◽  
Line P ◽  
Nanoparticle Shapes

Shape dependent uptake of glyco-gold nanoparticles (G-AuNPs) in a HeLa cell line.

scholarly journals SACCHAROMYCES CEREVISIAE-INDUCED APOPTOSIS OF MONOLAYER CERVICAL CANCER CELLS

2017 ◽  
Vol 10 (8) ◽  
pp. 63 ◽  
Author(s):  
Teena Rajan ◽  
Benluvankar V ◽  
Vincent S
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Hela Cell ◽  
Cell Line ◽  
Hela Cells ◽  
Prolonged Exposure ◽  
Chromatin Condensation ◽  
Yeast Cells ◽  
Apoptotic Cells ◽  
Hela Cell Line ◽  
Induction Of Apoptosis

  Objective: The present study was undertaken to examine the effect of phagocytosis of killed yeast on the induction of apoptosis in monolayer of HeLa cells.Methods: HeLa cell line was incubated with different doses (1000-7.8 μg/ml) of heat-killed Saccharomyces cerevisiae for 24, 48, and 72 hrs. The cytotoxicity against HeLa cell line during different exposure hours was screened by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-tetrazolium bromide assay. Induction of apoptosis was further confirmed by morphological and biochemical examination. Antiproliferative effect of yeast was examined under inverted microscope. Cell morphological changes were analyzed by fluorescent staining with propidium iodide.Results: The results showed that yeast induces cytotoxicity against HeLa cells in concentrations and during prolonged exposure periods. The viability of HeLa cells decreased from 85% to 45% during 72 hrs of treatment with 1000 μg/ml of yeast cells. The inhibitory concentration 50% of heat-killed yeast required to induce 50% inhibition of HeLa cells was 62.5 μg/ml. Apoptotic cells showed signs such as cell enlargement, membrane blebbing, and chromatin condensation. Furthermore, cell cycle analysis showed that S. cerevisiae treated HeLa cells and showed a typical apoptosis pattern of DNA content that reflected sub-G0 phase (corresponding to apoptotic cells).Conclusion: Results from the present work show that the heat-killed yeast has anticancer activity and it includes apoptosis of HeLa cells in vitro.

Identification and Expression Analysis of CD73 Inhibitors in Cervical Cancer

2020 ◽  
Vol 16 ◽  
Author(s):  
Jamshed Iqbal ◽  
Ayesha Basharat ◽  
Sehrish Bano ◽  
Syed Mobasher Ali Abid ◽  
Julie Pelletier ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cervical Cancer ◽  
Western Blot ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Apoptosis ◽  
Immunofluorescence Staining ◽  
Cell Cycle Analysis ◽  
Hela Cell Line ◽  
Cell Line Hela

Aims: The present study was conducted to examine the inhibitory effects of synthesized sulfonylhydrazones on the expression of CD73 (ecto-5′-NT). Background: CD73 (ecto-5′-NT) represents the most significant class of ecto-nucleotidases which are mainly responsible for dephosphorylation of adenosine monophosphate to adenosine. Inhibition of CD73 played an important role in the treatment of cancer, autoimmune disorders, precancerous syndromes, and some other diseases associated with CD73 activity. Objective: Keeping in view the significance of CD73 inhibitor in the treatment of cervical cancer, a series of sulfonylhydrazones (3a-3i) derivatives synthesized from 3-formylchromones were evaluated. Methods: All sulfonylhydrazones (3a-3i) were evaluated for their inhibitory activity towards CD73 (ecto-5′-NT) by the malachite green assay and their cytotoxic effect was investigated on HeLa cell line using MTT assay. Secondly, most potent compound was selected for cell apoptosis, immunofluorescence staining and cell cycle analysis. After that, CD73 mRNA and protein expression were analyzed by real-time PCR and Western blot. Results: Among all compounds, 3h, 3e, 3b, and 3c were found the most active against rat-ecto-5′-NT (CD73) enzyme with IC50 (µM) values of 0.70 ± 0.06 µM, 0.87 ± 0.05 µM, 0.39 ± 0.02 µM and 0.33 ± 0.03 µM, respectively. These derivatives were further evaluated for their cytotoxic potential against cancer cell line (HeLa). Compound 3h and 3c showed the cytotoxicity at IC50 value of 30.20 ± 3.11 µM and 86.02 ± 7.11 µM, respectively. Furthermore, compound 3h was selected for cell apoptosis, immunofluorescence staining and cell cycle analysis which showed promising apoptotic effect in HeLa cells. Additionally, compound 3h was further investigated for its effect on expression of CD73 using qRT-PCR and western blot. Conclusion: Among all synthesized compounds (3a-3i), Compound 3h (E)-N'-((6-ethyl-4-oxo-4H-chromen-3-yl) methylene)-4-methylbenzenesulfonohydrazide was identified as most potent compound. Additional expression studies conducted on HeLa cell line proved that this compound successfully decreased the expression level of CD73 and thus inhibiting the growth and proliferation of cancer cells.

Molecular events after antisense inhibition of hMSH2 in a HeLa cell line

1998 ◽  
Vol 418 (2-3) ◽  
pp. 61-71 ◽  
Author(s):  
Ying Qian ◽  
Yingnian Yu ◽  
Xingruo Cheng ◽  
Jianhong Luo ◽  
Haiyang Xie ◽  
...  
Keyword(s):  
Hela Cell ◽  
Cell Line ◽  
Antisense Inhibition ◽  
Hela Cell Line ◽  
Molecular Events

scholarly journals Synthesis, structural analysis, solution equilibria and biological activity of rhodium(iii) complexes with a quinquedentate polyaminopolycarboxylate

RSC Advances ◽  
2017 ◽  
Vol 7 (9) ◽  
pp. 5282-5296 ◽  
Author(s):  
Marija S. Jeremić ◽  
Hubert Wadepohl ◽  
Vesna V. Kojić ◽  
Dimitar S. Jakimov ◽  
Ratomir Jelić ◽  
...  
Keyword(s):  
Biological Activity ◽  
Structural Analysis ◽  
Antitumor Activity ◽  
Hela Cell ◽  
Cell Line ◽  
Hela Cell Line ◽  
Solution Equilibria ◽  
Line P

Two new Rh(iii)–ed3a complexes [Rh(ed3a)(OH2)]·H2O and Na[Rh(ed3a)Cl]·H2O have shown good antitumor activity, especially against HeLa cell line.

Exploring the potential of imines as antiprotozoan agents with focus on t. Brucei and p. Falciparum

2018 ◽  
Author(s):  
◽  
Kola Augustus Oluwafemi
Keyword(s):  
Hela Cell ◽  
Cell Line ◽  
Special Focus ◽  
Hela Cell Line ◽  
Free Energies ◽  
Imino Group ◽  
Design Synthesis ◽  
Lead Compounds ◽  
Pyridine Moiety ◽  
Nmr Study

This work focuses on the design, synthesis and evaluation of imine-containing heterocyclic and acyclic compounds with special focus on their bioactivity against parasitic protozoans (P. falciparum and T. brucei) - given the context of drug resistance in the treatment of malaria and Human African sleeping sickness and the fact that several bioactive organic compounds have been reported to possess the imino group. Starting from 2-aminopyridine, novel #-alkylated-5-bromo-7-azabenzimidazoles and substituted 5-bromo-1-(carbamoylmethy)-7-azabenzimidazole derivatives were prepared, and their bioactivity against parasitic protozoans was assessed. NMR spectra of the substituted 5- bromo-1-(carbamoylmethy)-7-azabenzimidazole derivatives exhibited rotational isomerism, and a dynamic NMR study was used in the estimation of the rate constants and the free- energies of activation for rotation. The free-energy differences between the two rotamers were determined and the more stable conformations were predicted. Novel 2-phenyl-7-azabenzimidazoles were also synthesised from 2-aminopyridine. A convenient method for the regioselective formylation of 2,3-diaminopyridines into 2-amino- 7-(benzylimino)pyridine analogues of 2-phenyl-7-azabenzimidazole was developed, and some of the resulting imino derivatives were hydrogenated to verify the importance of the imino moiety for bioactivity. The 2-phenyl-7-azabenzimidazoles and the 2-amino-7- (benzylimino)pyridine analogues were screened for their anti-protozoal activity and their cytotoxicity level was determined against the HeLa cell line. In order to validate the importance of the pyridine moiety, novel #-(phenyl)-2- hydroxybenzylimines, #-(benzyl)-2-hydroxybenzylimines and (±)-trans-1,2-bis[2- hydroxybenzylimino]cyclohexanes were also synthesized and screened for activity against the parasitic protozoans and for cytotoxicity against the HeLa cell line. The biological assay results indicated that these compounds are not significantly cytotoxic and a good number of them show potential as lead compounds for the development of new malaria and trypanosomiasis drugs.

331P Synergistic effects of ferula gummosa and radiotherapy to induce cytotoxicity and apoptosis in the hela cell line

2016 ◽  
Vol 27 (suppl_9) ◽  
Author(s):  
A. Fani-Pakdel ◽  
S.H. Forouzmand ◽  
S.H. Mousavi ◽  
V. Vazifedan ◽  
M. Nourbakhsh ◽  
...  
Keyword(s):  
Hela Cell ◽  
Cell Line ◽  
Synergistic Effects ◽  
Hela Cell Line

scholarly journals IN VITRO CYTOTOXICITY POTENTIAL OF ETHANOL EXTRACT OF SYZYGIUM SAMARANGENSE (Wt.)

2019 ◽  
Vol 6 (1) ◽  
pp. 30-32
Author(s):  
Poonkodi K ◽  
Mini R ◽  
Vimaladevi K ◽  
Prabhu V ◽  
Anusuya M ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Hela Cell ◽  
Cell Line ◽  
Ethanol Extract ◽  
In Vitro Cytotoxicity ◽  
Hela Cell Line ◽  
Phytochemical Screening ◽  
Ic50 Value ◽  
Dependent Activity

The present investigation is carried out to study the invitro cytotoxicity of ethanol extract of Syzygium samarangense leaves on HeLa cell line by using MTT assay. Ethanol extract of S. samarangense showed concentration dependent activity on HeLa cell line with IC50 value of 40.5 μg/ml which shows that ethanol extract of S. samarangense posses significant cytoxicity.Moreover the preliminary phytochemical screening showed the presence of fatty acids, alkaloids, flavonoids, terphenoids, saponins, tannins and steroids which are responsible for its cytotoxicity. There are only a few reports are available for cytotoxicity of ethanol extract of S. samarangense.

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