scholarly journals Overlapping distribution of two glycosyltransferases in the Golgi apparatus of HeLa cells.

1993 ◽  
Vol 120 (1) ◽  
pp. 5-13 ◽  
Author(s):  
T Nilsson ◽  
M Pypaert ◽  
M H Hoe ◽  
P Slusarewicz ◽  
E G Berger ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Hela Cell ◽  
Cell Line ◽  
Hela Cells ◽  
Hela Cell Line ◽  
Frozen Sections ◽  
Specific Antibodies ◽  
Trans Golgi Network ◽  
Endogenous Protein ◽  
Golgi Network

Thin, frozen sections of a HeLa cell line were double labeled with specific antibodies to localize the trans-Golgi enzyme, beta 1,4 galactosyltransferase (GalT) and the medial enzyme, N-acetylglucosaminyltransferase I (NAGT I). The latter was detected by generating a HeLa cell line stably expressing a myc-tagged version of the endogenous protein. GalT was found in the trans-cisterna and trans-Golgi network but, contrary to expectation, NAGT I was found both in the medial- and trans-cisternae, overlapping the distribution of GalT. About one third of the NAGT I and half of the GalT were found in the shared, trans-cisterna. These data show that the differences between cisternae are determined not by different sets of enzymes but by different mixtures.

Mapping the distribution of Golgi enzymes involved in the construction of complex oligosaccharides

1995 ◽  
Vol 108 (4) ◽  
pp. 1617-1627 ◽  
Author(s):  
C. Rabouille ◽  
N. Hui ◽  
F. Hunte ◽  
R. Kieckbusch ◽  
E.G. Berger ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Hela Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Hela Cells ◽  
Stable Expression ◽  
Hela Cell Line ◽  
Specific Antibodies ◽  
Hela Cell Lines ◽  
Golgi Network

The distribution of beta 1,2 N-acetylglucosaminyltransferase I (NAGT I), alpha 1,3-1,6 mannosidase II (Mann II), beta 1,4 galactosyltransferase (GalT), alpha 2,6 sialyltransferase (SialylT) was determined by immuno-labelling of cryo-sections from HeLa cell lines. Antibody labelling in the HeLa cell line was made possible by stable expression of epitope-tagged forms of these proteins or forms from species to which specific antibodies were available. NAGT I and Mann II had the same distribution occupying the medial and trans cisternae of the stack. GalT and SialylT also had the same distribution but they occupied the trans cisterna and the trans-Golgi network (TGN). These results generalise our earlier observations on the overlapping distribution of Golgi enzymes and show that each of the trans compartments of the Golgi apparatus in HeLa cells contains unique mixtures of those Golgi enzymes involved in the construction of complex, N-linked oligosaccharides.

scholarly journals Mitotic Golgi clusters are not tubular endosomes

1993 ◽  
Vol 104 (3) ◽  
pp. 811-818 ◽  
Author(s):  
M. Pypaert ◽  
T. Nilsson ◽  
E.G. Berger ◽  
G. Warren
Keyword(s):  
Hela Cell ◽  
Endocytic Pathway ◽  
Hela Cell Line ◽  
Frozen Sections ◽  
Qualitative And Quantitative ◽  
Quantitative Studies ◽  
Cell Biol ◽  
Endogenous Protein ◽  
Secondary Antibodies ◽  
Two Populations

HeLa cells were incubated with 15 nm BSA-gold for 1 or 2 hours to mark the endocytic pathway and mitotic cells were then isolated by shake-off. Thin, frozen sections were labelled with antibodies against two resident Golgi markers, beta-(1,4)-galactosyltransferase and N-acetylglucosaminyltransferase I. Detection of the latter was aided by the use of a HeLa cell line stably expressing a myc-tagged version of the endogenous protein. The secondary antibodies were coupled to either 5 or 10 nm gold so that the distribution of each of the three markers could be followed. Qualitative and quantitative studies showed that there were two populations of clusters, those described by us earlier and termed Golgi clusters (Lucocq et al. (1987) J. Cell Biol. 104, 865–874), containing either or both Golgi markers, and clusters of tubular endosomes containing BSA-gold. There was very little overlap showing that Golgi clusters cannot be tubular endosomes as concluded by Tooze and Hollinshead (1992) Eur. J. Cell Biol. 58, 228–242.

scholarly journals In vitro Anticancer Activity Effect of Extracellular Metabolites of Some Bacterial Species on HeLa Cell Line.

2020 ◽  
Keyword(s):  
Hela Cell ◽  
Anticancer Activity ◽  
Cell Line ◽  
Hela Cells ◽  
Bacterial Species ◽  
Hela Cell Line ◽  
Inhibitory Effects ◽  
Anticancer Effect ◽  
E Coli ◽  
Extracellular Metabolites

Background: Cancer is still one of the most serious problems that affect human health. Despite the intense efforts to develop treatments, effective agents are still not available. In some cases, conventional therapy could be harmful or fail because of emerging drug resistance. Therefore, the development of novel therapies against cancer is of utmost importance. Assessment of anticancer effects of bacterial metabolites on cancer cells may help in the process of finding new cheap, reliable, contentious and safe cancer therapy. Objective: To determine the anticancer effect of the extracellular metabolites of eight bacterial species on HeLa cell line. Methodology: Extracellular metabolites were prepared by isolating and culturing eight bacterial species (Escherichia coli, Staphylococcus aureus, Micrococcus, Pseudomonas aeruginosa, Lactic acid bacteria, Klebsiella, Proteus and E. coli with its phage) in liquid media. Tubes were incubated overnight and centrifuged. Supernatant was filtered and concentrated using Infra-Red concentrator. Different concentrations were prepared and their anticancer effect were evaluated using MTT cell proliferation assay. Results: Results showed variation among the eight bacteria concerning proliferation inhibition against HeLa cells in a time and concentration dependent manner. Pseudomonas and E. coli with its phage revealed considerable anticancer activity with 63% and 86% inhibitory effects (both at 1000 µg\ml) and IC50 of 301 and 1395 µg/dl at 24h respectively. While Proteus and Micrococcus showed low inhibitory effects and S. aureus enhanced the proliferation of HeLa cells at low concentrations. Conclusion: Among the tested bacteria, Pseudomonas and E. coli and its phage gave the best anticancer inhibitory effects against HeLa cells. Further studies using purified components of effective bacteria are recommended.

scholarly journals Investigation of the Dose-Enhancement Effects of Spherical and Rod-Shaped Gold Nanoparticles on the HeLa Cell Line

2020 ◽  
Vol 9 ◽  
pp. 1581
Author(s):  
Samad Amani ◽  
Alireza Mehdizadeh ◽  
Mohammad Mehdi Movahedi ◽  
Marzieh Keshavarz ◽  
Fereshteh Koosha
Keyword(s):  
Gold Nanoparticles ◽  
Hela Cell ◽  
Cell Line ◽  
Hela Cells ◽  
Current Treatment ◽  
X Rays ◽  
Hela Cell Line ◽  
X Ray ◽  
Cervical Cancer Cells ◽  
Gold Nanomaterials

Background: Cervical cancer cells are known as radioresistant cells. Current treatment methods have not improved the patients’ survival efficiently; thus, new therapeutic strategies are needed to enhance the efficacy of radiotherapy. Gold nanomaterials with different shapes and sizes have been explored as radiosensitizers. The present study compared the radiosensitizing effects of gold nanorods (AuNRs) with spherical gold nanoparticles (AuNPs) on the HeLa cell line irradiated with megavoltage X-rays. Materials and Methods: The cytotoxicity of AuNRs and AuNPs on HeLa cells in the presence and absence of 6-MV X-ray was investigated using the MTT assay. For this aim, HeLa cells were incubated with and AuNPs and AuNRs at various concentrations (5, 10, and 15 µg/mL) for 6 hours. Afterward, HeLa cells were irradiated with 6-MV X-ray at a single dose of 2 Gy. Results: The results showed that the addition of AuNRs and AuNPs could enhance the radiosensitivity of HeLa cells. Both AuNRs and AuNPs showed low toxicity on HeLa cells, while AuNRs were more toxic than AuNPs at the examined concentrations. Moreover, it was found that AuNRs could enhance the radiosensitivity of HeLa cells more than spherical-shaped AuNPs. Conclusion: This study revealed that the shape of nanoparticles is an effective factor when they are used as radiosensitizing agents during radiotherapy. [GMJ.2020;9:e1581]

scholarly journals Cytotoxic activity of Pinus cembra L. needle extract: A preliminary study on HeLa cell line

2021 ◽  
Vol 57 (2) ◽  
pp. 93-98
Author(s):  
Cristina LUNGU ◽  
◽  
Cosmin-Teodor MIHAI ◽  
Gabriela VOCHITA ◽  
Daniela GHERGHEL ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Hela Cell ◽  
Cell Line ◽  
Hela Cells ◽  
Pine Needles ◽  
Hela Cell Line ◽  
Pine Needle ◽  
Cytotoxic Effects ◽  
Antitumor Potential ◽  
Cembran Pine

The aim of this study was to investigate the cytotoxic effects of a hydromethanolic extract obtained from cembran pine needles in HeLa cell line. In this respect, the effects of needle extract on protein synthesis, viability, proliferation and cell cycle in HeLa cells were evaluated after 48 h treatment. Cembran pine needle extract dose-dependently decreased protein synthesis in HeLa cells causing 44.26% reduction in protein synthesis at 100µg/ml. At 25, 50 and 100µg/ml, it increased cell death in comparison with the control (20.99%, 21.49% and 23.63%, respectively vs. 9.83%). In addition, at 100µg/ml, cembran pine needle extract showed a remarkable antiproliferative effect whereas at 25 and 50µg/ml, it induced sub-G1 phase cells accumulation (11.68 ± 0.81% and 14.69 ± 0.56%, respectively in comparison with control, 6.03 ± 0.55%), an indicator of proapoptotic effects. Taken together, these results indicate that cembran pine needles are a source of compounds with antitumor potential which needs to be further investigated and exploited.

scholarly journals Comparison of Anticancer Effects of Hydroalcoholic Extracts of Camellia sinensis and Lepidium sativum L on HeLa Cell Line

2020 ◽  
Vol 13 (11) ◽  
Author(s):  
Somayeh Jahani ◽  
Zahra Heidari ◽  
Mehdi Azami ◽  
Bita Moudi
Keyword(s):  
Hela Cell ◽  
Growth Inhibition ◽  
Cell Line ◽  
Green Tea ◽  
Camellia Sinensis ◽  
Hela Cells ◽  
Green Tea Extract ◽  
Lepidium Sativum ◽  
Hela Cell Line ◽  
Tea Extract

Background: The antioxidative activity of green tea and garden cress extract is of interest in cancer. Objectives: The current study aimed at evaluating the effect of hydroalcoholic extracts of Lepidium sativum (cress) and Camellia sinensis (green tea) on the culture medium of the HeLa cell line. Methods: Dulbecco’s Modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) was used to culture HeLa cells, which was exposed to the different concentrations of green tea and cress extracts at 24 hours and 48 hours. Cell viability and apoptotic cells were quantified by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide or MTT assay and propidium iodide, respectively. Results: The highest percentage of growth inhibition (85%) was observed at 100 μg/mL of the green tea extract after 48-hour treatment. The percentage of growth inhibition at 24 h after treatment was 83% for green tea (P > 0.05). The high growth inhibition percentage of HeLa cells at 100 μg/mL of cress extract at 24 hours and 48 hours (49.8%) after treatment was 27.92% and 49.8%, respectively (P > 0.05). Additionally, the cell apoptosis assay indicated that green tea and cress extracts had toxic effects on the HeLa cells. This effect was highest at the concentration of 100 μg/mL and more evident in green tea. Conclusions: It can be concluded that green tea extract compared with cress had a more cytotoxic effect against cervical cancer.

scholarly journals Anticancer potentials of leaf, stem, and root extracts of Achyranthes aspera L.

2020 ◽  
Vol 12 (3) ◽  
pp. 546-555
Author(s):  
Nafisehsadat OMIDIANI ◽  
Kailas D. DATKHILE ◽  
Rajkumar B. BARMUKH
Keyword(s):  
Hela Cell ◽  
Ethyl Acetate ◽  
Cell Line ◽  
Hela Cells ◽  
Root Biomass ◽  
Acetone Extract ◽  
Hela Cell Line ◽  
Root Extracts ◽  
Achyranthes Aspera ◽  
Therapeutic Uses

Achyranthes aspera L. (Amaranthaceae), an herbaceous roadside weed in various parts of India, has several therapeutic uses, including the treatment against cancer. This investigation was undertaken to identify the bioactive compounds conferring cytotoxicity to the extracts from its biomass. The powdered leaf, stem, and root biomass was extracted separately in ethyl acetate, acetone, ethanol, and methanol. Each extract was tested against the HeLa cell line for the cytotoxicity, but the root-acetone extract was the most cytotoxic. This extract revealed eleven bands. The solution obtained from the ninth band (Rf = 0.87±0.06) exhibited more than 90% inhibition of HeLa cells. The LCMS analysis of this solution showed the presence of 37 compounds, out of which few compounds had been reported from different plants to possess cytotoxicity in various systems.

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.

scholarly journals Effects of elevated expression of inositol 1,4,5-trisphosphate 3-kinase B on Ca2+ homoeostasis in HeLa cells

2000 ◽  
Vol 352 (3) ◽  
pp. 709-715 ◽  
Author(s):  
Tom H. MILLARD ◽  
Peter J. CULLEN ◽  
George BANTING
Keyword(s):  
Hela Cell ◽  
Cell Line ◽  
Significant Role ◽  
Hela Cells ◽  
Second Messenger ◽  
Inducible Promoter ◽  
Hela Cell Line ◽  
Inositol 1,4,5 Trisphosphate ◽  
Elevated Expression ◽  
Stimulation Of

Ins(1,4,5)P3 3-kinase (IP3K) phosphorylates the Ca2+-mobilizing second messenger Ins(1,4,5)P3 to yield the putative second messenger Ins(1,3,4,5)P4. A HeLa cell line was established expressing the rat B isoform of IP3K under the control of an inducible promoter. The IP3KB-transfected cell line possessed 23-fold greater IP3K activity than untransfected cells after induction of IP3KB expression, but only 0.23-fold greater activity when IP3KB expression was not induced. Elevating IP3KB expression significantly reduced levels of Ins(1,4,5)P3 and increased levels of Ins(1,3,4,5)P4 after stimulation of cells with histamine, but had no effect on basal levels. Histamine- and ATP-evoked cytosolic Ca2+ responses were dramatically reduced upon elevation of IP3KB expression. On stimulation with a supramaximal dose of histamine, 67% of cells induced to express IP3KB gave no detectable elevation in cytosolic Ca2+, compared with 3% of uninduced cells. The quantity of Ca2+ within thapsigargin-sensitive and -insensitive stores was unaffected by elevation of IP3KB expression, as was capacitative Ca2+ entry. These data suggest that IP3KB may play a significant role in the regulation of Ins(1,4,5)P3 levels, and consequently in Ca2+ responses following stimulation of cells with Ins(1,4,5)P3-elevating agonists.

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.

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