scholarly journals Cytotoxic effects on cancerous and non-cancerous cells of trans-cinnamaldehyde, carvacrol, and eugenol

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Saurav Ranjitkar ◽  
Delong Zhang ◽  
Fei Sun ◽  
Saleh Salman ◽  
Wu He ◽  
...  
Keyword(s):  
Hela Cells ◽  
Normal Skin ◽  
Antioxidant Properties ◽  
Cell Types ◽  
Active Components ◽  
Cancer Treatments ◽  
Cytotoxic Effects ◽  
Reverse Transcription Quantitative Pcr ◽  
Apoptotic Genes ◽  
Cancerous Cells

AbstractEssential oils and their active components, referred here as plant derived antimicrobials (PDAs), have been used for their antimicrobial, anti-inflammatory and antioxidant properties. Many reports also document PDAs’ cytotoxic effects on cancerous cells, raising the hope that they could be used for cancer treatments. Due to the lack of specificity, we hypothesize that PDAs are cytotoxic to both cancerous and non-cancerous cells. Trans-cinnamaldehyde (TCA), carvacrol, and eugenol were assessed for their cytotoxicity on cancerous HeLa cells and normal skin fibroblasts (CCD-1123Sk, CCD) by MTT and LDH assays, flow cytometry, and reverse transcription quantitative PCR (RT-qPCR). After 24 h of treatment, carvacrol and TCA significantly decreased cell viability (by more than 50%) at 100 µg/ml, whereas eugenol was ineffective up to 400 µg/ml. Cell detachment and significantly increased apoptosis were observed with 100 µg/ml of TCA on both cell types. RT-qPCR for apoptotic genes (BCL2, CASP3 and CASP8) and necrosis genes (MLKL, RIPK1 and RIPK3) did not show significant differences between control and treated cells of both types, with the exception of eugenol-treated HeLa cells in which expression of BCL2, MLKL and RIPK1 was significantly higher than controls. Taken together, we conclude that the three PDAs studied here exhibited similar cytotoxic effects on both cancerous and non-cancerous cells.

scholarly journals Plant-derived Antimicrobials have Similar Cytotoxic Effects on Cancerous and Non-cancerous Cells

2021 ◽  
Author(s):  
Saurav Ranjitkar ◽  
Delong Zhang ◽  
Fei Sun ◽  
Saleh Salman ◽  
Wu He ◽  
...  
Keyword(s):  
Hela Cells ◽  
Normal Skin ◽  
Antioxidant Properties ◽  
Cell Types ◽  
Active Components ◽  
Cancer Treatments ◽  
Cytotoxic Effects ◽  
Reverse Transcription Quantitative Pcr ◽  
Apoptotic Genes ◽  
Cancerous Cells

Abstract Essential oils and their active components, referred here as plant derived antimicrobials (PDAs), have been used for their antimicrobial, anti-inflammatory and antioxidant properties. Many reports also document PDAs’ cytotoxic effects on cancerous cells, raising the hope that they could be used for cancer treatments. Due to the lack of specificity, we hypothesize that PDAs are cytotoxic to both cancerous and non-cancerous cells. Trans-cinnamaldehyde (TCA), carvacrol, and eugenol were assessed for their cytotoxicity on cancerous HeLa cells and normal skin fibroblasts (CCD-1123Sk, CCD) by MTT and LDH assays, flow cytometry, and reverse transcription quantitative PCR (RT-qPCR). After 24 hours of treatment, carvacrol and TCA significantly decreased cell viability (by more than 50%) at 100 µg/ml, whereas eugenol was ineffective up to 400 µg/ml. Cell detachment and significantly increased apoptosis were observed with 100 µg/ml of TCA on both cell types. RT-qPCR for apoptotic genes (Bcl2, Casp3 and Casp8) and necrosis genes (Mlkl, Ripk1 and Ripk3) did not show significant differences between control and treated cells of both types, with the exception of eugenol-treated HeLa cells in which expression of Bcl2, Mlkl and Ripk1 was significantly higher than controls. Taken together, we conclude that the three PDAs studied here exhibited similar cytotoxic effects on both cancerous and non-cancerous cells.

scholarly journals Satureja subspicata and S. horvatii Extracts Induce Overexpression of the BCl-2 Family of Anti-apoptotic Genes and Reduce Micronuclei Frequency in Mice

2018 ◽  
Vol 13 (6) ◽  
pp. 1934578X1801300
Author(s):  
Jasmina Čakar ◽  
Naida Kadrić Lojo ◽  
Anja Haverić ◽  
Maida Hadžić ◽  
Lejla Lasić ◽  
...  
Keyword(s):  
Human Lymphocyte ◽  
Micronucleus Assay ◽  
Balkan Peninsula ◽  
Lymphocyte Culture ◽  
Cytotoxic Activities ◽  
Cytotoxic Effects ◽  
Apoptotic Genes ◽  
Apoptotic Activity

Satureja subspicata and S. horvatii are endemic species of the Balkan Peninsula and often used in traditional medicine in Bosnia and Herzegovina to treat different health conditions. We aimed to analyze the unevaluated apoptotic, genotoxic and cytotoxic effects of two Satureja species, as well as their content of phenolics that are mainly responsible for the plant's biological activity. Apoptotic and geno/cytotoxic activities of S. subspicata and S. horvatii were investigated in vitro in human lymphocyte culture and in vivo in mice. The content of the main phenolics in plant extracts was determined by ultra-high pressure liquid chromatography-MS-MS (UHPLC–MS/MS). Genotoxic and cytotoxic activities of Satureja extracts were evaluated in vitro by applying a cytokinesis-block micronucleus cytome assay in human lymphocyte culture and in vivo applying a mice reticulocytes micronucleus assay. SALSA RT-MLPA R011-C1 apoptosis assay was used for measuring the relative expression of 44 genes associated with the regulation of the apoptotic pathways in human lymphocyte cultures treated with different concentrations of two Satureja extracts. The first analysis of phenolic compounds in S. horvatii and S. subspicata determined by an UHPLC-MS/MS method revealed high levels of rosmarinic and caffeic acids. Minor genotoxic potential was determined in relation to the tested concentrations while no cytostatic and cytotoxic effects were revealed in vitro. However, when applied in concentrations of 200 mg/kg per os, aqueous extracts of two Satureja species significantly decreased frequency of reticulocytes micronuclei in treated mice against controls. Extracts of S. subspicata and S. horvatii in concentrations of 0.2 mg/mL, regardless of solvent used, downregulated pro-apoptotic and upregulated anti-apoptotic genes, showing anti-apoptotic activity. Our results indicate that the registered anti-genotoxic and anti-apoptotic activity is most likely related to the high level of phenolic acids (particularly rosmarinic and caffeic) in the tested extracts.

Interaction of Candida albicans with genital mucosa: effect of sex hormones on adherence of yeasts in vitro

1988 ◽  
Vol 34 (3) ◽  
pp. 224-228 ◽  
Author(s):  
Aliza Kalo ◽  
Esther Segal
Keyword(s):  
Candida Albicans ◽  
Hela Cell ◽  
Sex Hormones ◽  
Hela Cells ◽  
Cell Types ◽  
Vaginal Candidiasis ◽  
Genital Mucosa ◽  
Hela Cell Lines ◽  
I Cells

Findings from our previous studies revealed a correlation between the level of adherence in vitro of Candida albicans to human exfoliated vaginal epithelial cells (VEC) and the hormonal status of the cell donors. In the present study we investigated the effect of the sex hormones estradiol, estriol, progesterone, and testosterone on the binding of the yeasts to HeLa cell lines and VEC in vitro. Monolayers of HeLa cells were exposed to the hormones and yeasts under controlled conditions. The number of adherent yeasts per square millimetre of HeLa cell monolayers and the percentage of VEC with adherent yeasts was estimated by microscopic counts. The results showed that the tested sex hormones affected at various degrees the adhesion of yeasts to HeLa cells or VEC. Progesterone had the most marked effect, leading to a significant increase in the number of adherent yeasts to HeLa cells or in the percentage of adhesion of VEC. In addition, VEC were separated on Percoll gradients into the two cell types: superficial (S) and intermediate (I), cell types which appear physiologically under increased serum levels of estradiol or progesterone, respectively. Adhesion assays with the separated cell populations revealed an increased binding capacity of the I cells. The finding that progesterone increased the adherence of yeasts to genital mucosa and that VEC of the I type have a higher capacity to adhere the yeasts is compatible with our previous observation that increased numbers of I cells, appearing under high level of progesterone, are found in situations known to have predisposition to vaginal candidiasis. Thus, our data point to a possible involvement of the hormone progesterone in the adherence of C. albicans to genital epithelium.

scholarly journals Evaluation of Cytotoxic Effects of Dichloromethane Extract of Guduchi (Tinospora cordifolia Miers ex Hook F & THOMS) on Cultured HeLa Cells

2006 ◽  
Vol 3 (2) ◽  
pp. 267-272 ◽  
Author(s):  
Ganesh Chandra Jagetia ◽  
Shaival Kamalaksha Rao
Keyword(s):  
Lipid Peroxidation ◽  
Lactate Dehydrogenase ◽  
Hela Cells ◽  
Cytotoxic Effect ◽  
Tinospora Cordifolia ◽  
Cytotoxic Effects ◽  
Optimal Duration ◽  
Ldh Release ◽  
Gst Activity

Extracts ofTinospora cordifolia(TCE) have been shown to possess anti-tumor properties, but the mechanism of the anti-tumor function of TCE is poorly understood. This investigation elucidates the possible mechanism underlying the cytotoxic effects of dichlormethane extracts of TCE, after selecting optimal duration and concentration for treatment. HeLa cells were exposed to various concentrations of TCE, which has resulted in a concentration-dependent decline in the clonogenicity, glutathione-S-transferase (GST) activity and a concentration-dependent increase in lipid peroxidation (TBARS) with a peak at 4 h and lactate dehydrogenase (LDH) release with a peak at 2 h. Our results suggest that the cytotoxic effect of TCE may be due to lipid peroxidation and release of LDH and decline in GST.

scholarly journals Microinjection of Francisella tularensis and Listeria monocytogenes Reveals the Importance of Bacterial and Host Factors for Successful Replication

2015 ◽  
Vol 83 (8) ◽  
pp. 3233-3242 ◽  
Author(s):  
Lena Meyer ◽  
Jeanette E. Bröms ◽  
Xijia Liu ◽  
Martin E. Rottenberg ◽  
Anders Sjöstedt
Keyword(s):  
Listeria Monocytogenes ◽  
Francisella Tularensis ◽  
Hela Cells ◽  
Cell Types ◽  
Metabolic Adaptation ◽  
Content Type ◽  
Bacterial Uptake ◽  
J774 Cells ◽  
Cell Cytosol ◽  
Successful Replication

Certain intracellular bacteria use the host cell cytosol as the replicative niche. Although it has been hypothesized that the successful exploitation of this compartment requires a unique metabolic adaptation, supportive evidence is lacking. ForFrancisella tularensis, many genes of theFrancisellapathogenicity island (FPI) are essential for intracellular growth, and therefore, FPI mutants are useful tools for understanding the prerequisites of intracytosolic replication. We compared the growth of bacteria taken up by phagocytic or nonphagocytic cells with that of bacteria microinjected directly into the host cytosol, using the live vaccine strain (LVS) ofF. tularensis; five selected FPI mutants thereof, i.e., ΔiglA, ΔiglÇ ΔiglG, ΔiglI, and ΔpdpEstrains; andListeria monocytogenes. After uptake in bone marrow-derived macrophages (BMDM), ASC−/−BMDM, MyD88−/−BMDM, J774 cells, or HeLa cells, LVS, ΔpdpEand ΔiglGmutants, andL. monocytogenesreplicated efficiently in all five cell types, whereas the ΔiglAand ΔiglCmutants showed no replication. After microinjection, all 7 strains showed effective replication in J774 macrophages, ASC−/−BMDM, and HeLa cells. In contrast to the rapid replication in other cell types,L. monocytogenesshowed no replication in MyD88−/−BMDM and LVS showed no replication in either BMDM or MyD88−/−BMDM after microinjection. Our data suggest that the mechanisms of bacterial uptake as well as the permissiveness of the cytosolic compartmentper seare important factors for the intracytosolic replication. Notably, none of the investigated FPI proteins was found to be essential for intracytosolic replication after microinjection.

Differential expression of the human gonadotropin alpha gene in ectopic and eutopic cells

1985 ◽  
Vol 5 (11) ◽  
pp. 3157-3167
Author(s):  
R B Darnell ◽  
I Boime
Keyword(s):  
Hela Cells ◽  
Cell Types ◽  
Base Pairs ◽  
Gene Encoding ◽  
Basal Expression ◽  
Bacterial Enzyme ◽  
Specific Induction ◽  
Alpha Gene ◽  
Cat Gene ◽  
Jar Cells

We have analyzed the regulation of the alpha gonadotropin gene in eutopic placental cells and ectopic tumor cells by constructing a series of plasmid vectors containing alpha genomic 5' flanking DNA placed upstream of the gene encoding the bacterial enzyme chloramphenicol acetyltransferase (CAT). These plasmid DNAs were transfected into a eutopic (JAr) and an ectopic (HeLa) cell line. Both cell types expressed the CAT gene from plasmid constructs containing as much as 1,500 base pairs (bp) and as little as 140 bp of alpha 5' flanking DNA; JAr cells were considerably more efficient than HeLa cells. Ectopic and eutopic cells differed qualitatively in their expression from these alpha-CAT constructs when cells were treated with cAMP or butyrate. Butyrate induced alpha expression in HeLa cells but not in JAr cells, while cAMP induced expression in JAr cells. These results are consistent with and extend previous observations suggesting that there are cell-specific differences in the regulation of alpha gene expression in ectopic and eutopic cells. However, by using deletion constructs of the alpha-CAT gene, we found that the basal expression and cell-specific induction of the alpha gene in ectopic and eutopic cells were dependent on the same 140 bp of alpha 5' flanking DNA. These 140 bp were sequenced and found to contain a 9-bp stretch of DNA homologous with the consensus viral enhancer sequence. Such features of alpha expression common to both ectopic and eutopic cells may be involved in the coordinate expression of the alpha gene and the tumorigenic phenotype observed in each cell type.

Different cis-acting DNA elements control expression of the human apolipoprotein AI gene in different cell types

1988 ◽  
Vol 8 (2) ◽  
pp. 605-614
Author(s):  
K N Sastry ◽  
U Seedorf ◽  
S K Karathanasis
Keyword(s):  
Hela Cells ◽  
Hepatoma Cell ◽  
Plasma Lipid ◽  
Cell Types ◽  
Apolipoprotein Ai ◽  
Transcriptional Enhancer ◽  
Regulation Of Expression ◽  
Cis Acting ◽  
Human Apolipoprotein ◽  
Mammalian Liver

In mammals, the gene coding for apolipoprotein AI (apoAI), a protein of the plasma lipid transport system, is expressed only in the liver and the intestine. A series of plasmids containing various lengths of sequences flanking the 5' end of the human apoAI gene were constructed and assayed for transient expression after introduction into cultured human hepatoma (HepG2), colon carcinoma (Caco-2), and epithelial (HeLa) cells. The results showed that while most of these constructs are expressed in HepG2 and Caco-2 cells, none of them is expressed in HeLa cells. In addition, the results indicated that a DNA segment located between nucleotides -256 and -41 upstream from the transcription start site of this gene is necessary and sufficient for maximal levels of expression in HepG2 but not in Caco-2 cells, while a DNA segment located between nucleotides -2052 and -192 is required for maximal levels of expression in Caco-2 cells. Moreover, it was shown that the -256 to -41 DNA segment functions as a hepatoma cell-specific transcriptional enhancer with both homologous and heterologous promoters. These results indicate that different cis- and possibly trans-acting factors are involved in the establishment and subsequent regulation of expression of the apoAI gene in the mammalian liver and intestine.

scholarly journals Hepatitis C Virus Core Protein Enhances NF-κB Signal Pathway Triggering by Lymphotoxin-β Receptor Ligand and Tumor Necrosis Factor Alpha

1999 ◽  
Vol 73 (2) ◽  
pp. 1672-1681 ◽  
Author(s):  
Li-Ru You ◽  
Chun-Ming Chen ◽  
Yan-Hwa Wu Lee
Keyword(s):  
Tumor Necrosis Factor ◽  
Hela Cells ◽  
Tumor Necrosis ◽  
Core Protein ◽  
Cell Types ◽  
Necrosis Factor Alpha ◽  
Hcv Core Protein ◽  
Tnf Α ◽  
Β Receptor ◽  
Necrosis Factor

ABSTRACT Our previous study indicated that the core protein of hepatitis C virus (HCV) can associate with tumor necrosis factor receptor (TNFR)-related lymphotoxin-β receptor (LT-βR) and that this protein-protein interaction plays a modulatory effect on the cytolytic activity of recombinant form LT-βR ligand (LT-α1β2) but not tumor necrosis factor alpha (TNF-α) in certain cell types. Since both TNF-α/TNFR and LT-α1β2/LT-βR are also engaged in transcriptional activator NF-κB activation or c-Jun N-terminal kinase (JNK) activation, the biological effects of the HCV core protein on these regards were elucidated in this study. As demonstrated by the electrophoretic mobility shift assay, the expression of HCV core protein prolonged or enhanced the TNF-α or LT-α1β2-induced NF-κB DNA-binding activity in HuH-7 and HeLa cells. The presence of HCV core protein in HeLa or HuH-7 cells with or without cytokine treatment also enhanced the NF-κB-dependent reporter plasmid activity, and this effect was more strongly seen with HuH-7 cells than with HeLa cells. Western blot analysis suggested that this modulation of the NF-κB activity by the HCV core protein was in part due to elevated or prolonged nuclear retention of p50 or p65 species of NF-κB in core protein-producing cells with or without cytokine treatment. Furthermore, the HCV core protein enhanced or prolonged the IκB-β degradation triggering by TNF-α or LT-α1β2 both in HeLa and HuH-7 cells. In contrast to that of IκB-β, the increased degradation of IκB-α occurred only in LT-α1β2-treated core-producing HeLa cells and not in TNF-α-treated cells. Therefore, the HCV core protein plays a modulatory effect on NF-κB activation triggering by both cytokines, though the mechanism of NF-κB activation, in particular the regulation of IκB degradation, is rather cell line and cytokine specific. Studies also suggested that the HCV core protein had no effect on TNF-α-stimulated JNK activity in both HeLa and HuH-7 cells. These findings, together with our previous study, strongly suggest that among three signaling pathways triggered by the TNF-α-related cytokines, the HCV core protein potentiates NF-κB activation in most cell types, which in turn may contribute to the chronically activated, persistent state of HCV-infected cells.

scholarly journals Characterisation of GLUT4 trafficking in HeLa cells: comparable kinetics and orthologous trafficking mechanisms to 3T3-L1 adipocytes

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8751 ◽  
Author(s):  
Silke Morris ◽  
Niall D. Geoghegan ◽  
Jessica B.A. Sadler ◽  
Anna M. Koester ◽  
Hannah L. Black ◽  
...  
Keyword(s):  
Cell Surface ◽  
Hela Cells ◽  
Glucose Transporter ◽  
Characteristic Property ◽  
Cell Types ◽  
Model System ◽  
Human Model ◽  
Small Scale ◽  
Glucose Transporter Glut4 ◽  
Surface Fusion

Insulin-stimulated glucose transport is a characteristic property of adipocytes and muscle cells and involves the regulated delivery of glucose transporter (GLUT4)-containing vesicles from intracellular stores to the cell surface. Fusion of these vesicles results in increased numbers of GLUT4 molecules at the cell surface. In an attempt to overcome some of the limitations associated with both primary and cultured adipocytes, we expressed an epitope- and GFP-tagged version of GLUT4 (HA–GLUT4–GFP) in HeLa cells. Here we report the characterisation of this system compared to 3T3-L1 adipocytes. We show that insulin promotes translocation of HA–GLUT4–GFP to the surface of both cell types with similar kinetics using orthologous trafficking machinery. While the magnitude of the insulin-stimulated translocation of GLUT4 is smaller than mouse 3T3-L1 adipocytes, HeLa cells offer a useful, experimentally tractable, human model system. Here, we exemplify their utility through a small-scale siRNA screen to identify GOSR1 and YKT6 as potential novel regulators of GLUT4 trafficking in human cells.

scholarly journals Effects of inducing or inhibiting apoptosis on Sindbis virus replication in mosquito cells

2008 ◽  
Vol 89 (11) ◽  
pp. 2651-2661 ◽  
Author(s):  
Hua Wang ◽  
Carol D. Blair ◽  
Ken E. Olson ◽  
Rollie J. Clem
Keyword(s):  
Virus Replication ◽  
Persistent Infection ◽  
Sindbis Virus ◽  
Actinomycin D ◽  
Virus Production ◽  
Cell Types ◽  
Lytic Infection ◽  
Mosquito Cells ◽  
Infected Cells ◽  
Apoptotic Genes

Sindbis virus (SINV) is a mosquito-borne virus in the genus Alphavirus, family Togaviridae. Like most alphaviruses, SINVs exhibit lytic infection (apoptosis) in many mammalian cell types, but are generally thought to cause persistent infection with only moderate cytopathic effects in mosquito cells. However, there have been several reports of apoptotic-like cell death in mosquitoes infected with alphaviruses or flaviviruses. Given that apoptosis has been shown to be an antiviral response in other systems, we have constructed recombinant SINVs that express either pro-apoptotic or anti-apoptotic genes in order to test the effects of inducing or inhibiting apoptosis on SINV replication in mosquito cells. Recombinant SINVs expressing the pro-apoptotic genes reaper (rpr) from Drosophila or michelob_x (mx) from Aedes aegypti caused extensive apoptosis in cells from the mosquito cell line C6/36, thus changing the normal persistent infection observed with SINV to a lytic infection. Although the infected cells underwent apoptosis, high levels of virus replication were still observed during the initial infection. However, virus production subsequently decreased compared with persistently infected cells, which continued to produce high levels of virus over the next several days. Infection of C6/36 cells with SINV expressing the baculovirus caspase inhibitor P35 inhibited actinomycin D-induced caspase activity and protected infected cells from actinomycin D-induced apoptosis, but had no observable effect on virus replication. This study is the first to test directly whether inducing or inhibiting apoptosis affects arbovirus replication in mosquito cells.

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