scholarly journals Cytotoxicity and Apoptosis Induced by Chenopodium ambrosioides L. Essential Oil in Human Normal Liver Cell Line L02 via the Endogenous Mitochondrial Pathway Rather Than the Endoplasmic Reticulum Stress

2021 ◽  
Vol 18 (14) ◽  
pp. 7469
Author(s):  
Xiao-Ying Wang ◽  
Jun-Mei Hao ◽  
Qiu-Rong Ren ◽  
Hai-Ying Li ◽  
Jing-Song Wu ◽  
...  
Keyword(s):  
Essential Oil ◽  
Normal Liver ◽  
Mitochondrial Pathway ◽  
S Phase ◽  
Fluorescence Microscope ◽  
Apoptotic Cells ◽  
Chenopodium Ambrosioides ◽  
Normal Liver Cell ◽  
L02 Cells ◽  
Normal Liver Cell Line

Chenopodium ambrosioides L. (C. ambrosioides) has been used as dietary condiments and as traditional medicine in South America. The oil of Chenopodium ambrosioides L. (C. ambrosioides) can be used as a natural antioxidant in food processing. It also has analgesic, sedating, and deworming effects, and can be used along with the whole plant for its medical effects: decongestion, as an insecticide, and to offer menstruation pain relief. This study was conducted to investigate the cytotoxicity and apoptosis effects of an essential oil from C. ambrosioides in vitro. The cytotoxicity evaluation of the essential oil from C. ambrosioides on human normal liver cell line L02 was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. AO/EB dual fluorescent staining assay and Annexin V-FITC were used for apoptosis analysis. The changes in mitochondrial membrane potential (MMP) were analyzed with 5,5,6,6′-tetrachloro-1,1,3,3,-tetraethyl-imidacarbocyanine iodide (JC-1) dye under a fluorescence microscope. The level of apoptosis related protein expression was quantified by Western blot. The L02 cells were treated with the essential oil from C. ambrosioides at 24, 48, and 72 h, and the IC50 values were 65.45, 58.03, and 35.47 μg/mL, respectively. The AO/EB staining showed that viable apoptotic cells, non-viable apoptotic cells, and non-viable non-apoptotic cells appeared among the L02 cells under the fluorescence microscope. Cell cycle arrest at the S phase and cell apoptosis increased through flow cytometry in the L02 cells treated with the essential oil. MMP decreased in a concentration-dependent manner, as seen through JC-1 staining under the fluorescence microscope. In the L02 cells as shown by Western blot and qPCR, the amount of the apoptosis-related proteins and the mRNA expression levels of cytochrome C, Bax, Caspase-9, and Caspase-3 increased, Bcl-2 decreased, and Caspase-12, which is expressed in the endoplasmic reticulum, showed no obvious changes in protein amount or mRNA expression level. The essential oil form C. ambrosioides had a cytotoxic effect on L02 cells. It could inhibit L02 cell proliferation, arrest the cell cycle at the S phase, and induce L02 cell apoptosis through the endogenous mitochondrial pathway.

scholarly journals The cytotoxicity effect of Metalloprotease produced and isolated from Aeromonas spp.

2018 ◽  
Vol 42 (1) ◽  
pp. 41-45
Author(s):  
Muhannad M. Abdallah
Keyword(s):  
Aeromonas Hydrophila ◽  
Cytotoxic Effect ◽  
Hepatocellular Cancer ◽  
Normal Liver ◽  
Gel Chromatography ◽  
Chain Reaction ◽  
Normal Liver Cell ◽  
Ammonium Sulphate Precipitation ◽  
Polymerase Chain ◽  
Normal Liver Cell Line

     One hundred and fifty stool sampling were collected from clinical sources responsible of causing diarrhea in adults as well as children. The identification of Aeromonas hydrophila isolates depended on common methods of identification dependent on biochemical characteristics and culture, then vitek2 compact system was used. Eight Aeromonas hydrophila isolates were gained and revealed various productivity of metalloprotease; the isolate number 8 was the maximum effective in metalloprotease production. The eight isolates were examined with Polymerase Chain Reaction to prove enzyme gene presence, the results revealed that all isolates were positive for ahMP genes; metalloprotease was completely purified via a number of steps, which included ammonium sulphate precipitation, dialysis, ion exchange and gel chromatography. Cytotoxicity effect of metalloprotease studies on cancer and normal cell lines, The results showed the purified metalloprotease as effective cytotoxic effect on liver hepatocellular cancer cells (HepG2) compared with no effect on normal liver cell line (WRL-68) indicating less cytotoxic effect.

scholarly journals Novel Hybrid Compounds Containing Benzofuroxan and Aminothiazole Scaffolds: Synthesis and Evaluation of Their Anticancer Activity

2021 ◽  
Vol 22 (14) ◽  
pp. 7497
Author(s):  
Elena Chugunova ◽  
Gabriele Micheletti ◽  
Dario Telese ◽  
Carla Boga ◽  
Daut Islamov ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Normal Liver ◽  
Mitochondrial Pathway ◽  
Reaction Pathways ◽  
Aromatic Nucleophilic Substitution ◽  
X Ray ◽  
Hybrid Compounds ◽  
Aromatic Nucleophilic Substitution Reaction ◽  
Further Development

A series of novel hybrid compounds containing benzofuroxan and 2-aminothiazole moieties are synthesized via aromatic nucleophilic substitution reaction. Possible reaction pathways have been considered quantum-chemically, which allowed us to suggest the most probable products. The quantum chemical results have been proved by X-ray data on one compound belonging to the synthesized series. It was shown that the introduction of substituents to both the thiazole and amine moieties of the compounds under study strongly influences their UV/Vis spectra. Initial substances and obtained hybrid compounds have been tested in vitro as anticancer agents. Target compounds showed selectivity towards M-HeLa tumor cell lines and were found to be more active than starting benzofuroxan and aminothiazoles. Furthermore, they are considerably less toxic to normal liver cells compared to Тamoxifen. The mechanism of action of the studied compounds can be associated with the induction of apoptosis, which proceeds along the mitochondrial pathway. Thus, new hybrids of benzofuroxan are promising candidates for further development as anticancer agents.

scholarly journals Selective Anti-Cancer Effects of Plasma-Activated Medium and Its High Efficacy with Cisplatin on Hepatocellular Carcinoma with Cancer Stem Cell Characteristics

2021 ◽  
Vol 22 (8) ◽  
pp. 3956
Author(s):  
Yan Li ◽  
Tianyu Tang ◽  
Hae June Lee ◽  
Kiwon Song
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Death ◽  
Cell Lines ◽  
Primary Liver Cancer ◽  
Atmospheric Pressure Plasma ◽  
Normal Liver ◽  
Normal Liver Cell ◽  
Ample Evidence ◽  
Huh7 Cells ◽  
Anti Cancer

Hepatocellular carcinoma (HCC) is a major histological subtype of primary liver cancer. Ample evidence suggests that the pathological properties of HCC originate from hepatic cancer stem cells (CSCs), which are responsible for carcinogenesis, recurrence, and drug resistance. Cold atmospheric-pressure plasma (CAP) and plasma-activated medium (PAM) induce apoptosis in cancer cells and represent novel and powerful anti-cancer agents. This study aimed to determine the anti-cancer effect of CAP and PAM in HCC cell lines with CSC characteristics. We showed that the air-based CAP and PAM selectively induced cell death in Hep3B and Huh7 cells with CSC characteristics, but not in the normal liver cell line, MIHA. We observed both caspase-dependent and -independent cell death in the PAM-treated HCC cell lines. Moreover, we determined whether combinatorial PAM therapy with various anti-cancer agents have an additive effect on cell death in Huh7. We found that PAM highly increased the efficacy of the chemotherapeutic agent, cisplatin, while enhanced the anti-cancer effect of doxorubicin and the targeted-therapy drugs, trametinib and sorafenib to a lesser extent. These findings support the application of CAP and PAM as anti-cancer agents to induce selective cell death in cancers containing CSCs, suggesting that the combinatorial use of PAM and some specific anti-cancer agents is complemented mechanistically.

Mitomycin Induced Apoptosis in Human Leukemia K562 Cells

2012 ◽  
Vol 599 ◽  
pp. 71-75
Author(s):  
Shu Li Shao ◽  
Bin Zhao ◽  
Wei Wei Zhang ◽  
Wei Zhao ◽  
Guang Hui Wu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Gel Electrophoresis ◽  
Light Microscope ◽  
Previous Experiment ◽  
K562 Cells ◽  
S Phase ◽  
Fluorescence Microscope ◽  
Human Leukemia ◽  
Induced Apoptosis ◽  
Human Leukemia K562 Cells

Objective: The research aimed to study the effects of mitomycin on human leukemic K562 cells, and to explore the mechanism of mitomycin induced apoptosis．In order to provide previous experiment basis for mitomycin applying clinical treatments Methods: The multiplication and apoptosis status of K562 cells treated different time by different concentration mitomycin were observed by light microscope, fluorescence microscope, TEM, agrose gel electrophoresis of DNA and flow cytometry. Results: The results showed that mitomycin could induce K562 cells apoptosis, and the best concentration was 12.5μg/ml for 48 h. The optimal concentration of apoptosis induced by apoptosis rate is （28.8±1.04）% （P<0.01）. Mitomycin could affect the S phase among cellular multiplication, cell could be blocked by mitomycin and then apoptosis in this phase. Conclusions: Mitomycin can induce the apoptosis of human leukemic K562 cells. It is of great significance to guide clinical medication.

scholarly journals C/EBPα Inhibits Cell Growth via Direct Repression of E2F-DP-Mediated Transcription

2000 ◽  
Vol 20 (16) ◽  
pp. 5986-5997 ◽  
Author(s):  
Beatrix A. Slomiany ◽  
Kenneth L. D'Arigo ◽  
Margaret M. Kelly ◽  
David T. Kurtz
Keyword(s):  
Cell Growth ◽  
Ectopic Expression ◽  
Simian Virus 40 ◽  
Normal Liver ◽  
Simian Virus ◽  
Binding Activity ◽  
T Antigen ◽  
S Phase ◽  
3T3 Cells ◽  
L Cells

ABSTRACT Using an inducible transcription system which allows the regulated expression of C/EBP isoforms in tissue culture cells, we have found that the ectopic expression of C/EBPα, at a level comparable to that found in normal liver tissue, has a pronounced antimitogenic effect in mouse L cells and NIH 3T3 cells. The inhibition of cell division by C/EBPα in mouse cells cannot be reversed by simian virus 40 T antigen, by oncogenic ras, or by adenovirus E1a protein. When expressed in thymidine kinase-deficient L cells or 3T3 cells, C/EBPα is detected in a protein complex which binds to the E2F binding sites found in the promoters of the genes for E2F-1 and dihydrofolate reductase (DHFR). Bacterially expressed C/EBPα has no affinity for these E2F sites, but when recombinant C/EBPα is added to nuclear extracts from mouse fibroblasts, a new E2F binding activity appears, which contains the C/EBPα protein. Using an E2F-DP1-responsive promoter linked to a reporter gene, it can be shown that C/EBPα directly inhibits the induction of this promoter by E2F-DP1 in transient-transfection assays. Furthermore, C/EBPα can be shown to inhibit the S-phase induction of the E2F and DHFR promoters in permanent cell lines. These findings delineate a straightforward mechanism for C/EBPα-mediated cell growth arrest through repression of E2F-DP-mediated S-phase transcription.

scholarly journals Total Phenols, Identification of Active Compounds and Anticancer Activity of Salvia judaica Boiss against the breast Cancer Cell MDA-231

2021 ◽  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Essential Oil ◽  
Anticancer Activity ◽  
Breast Cancer Cell ◽  
Annexin V ◽  
Ethanol Extract ◽  
G1 Phase ◽  
Apoptotic Cells ◽  
Total Phenols

Salvia judaica is an annual herb from genus Salvia L.; the largest genera of Lamiaceae. It’s a medicinal plant prominent in pharmaceutical applications in many countries around the world. This study aimed to explore bioactive compounds likely to be responsible for the plant anticancer activity, and evaluate anticancer effects, after determining the total content of phenols in the ethanol extract and essential oil in this species. Ethanol extract (EE) and essential oil (EO) were prepared from dried aerial parts (leaves and the flower). GC-MS analysis of EO showed the presence of/43/ effective compounds in varying proportions, the major compounds were sesquiterpenes like delta-cadinene, alpha-Gurjunene, beta-humulene, and alpha-caryophyllene. This is the first study revealed that S.judaica is so rich in phenols which proceeded S.officinalis, noting the superiority of the EE over the EO samples in the total phenols. Anticancer properties of EE and EO of S. judaica against MDA-231 breast cancer cell line were studied -for the first time - by cell cycle analysis and Annexin V/PI apoptosis assay using Flow cytometry technique. Cells were treated with EE (0.001, 0.01, 0.02, 0.1mg/ml) and EO (0.005, 0.01, 0.02, 0.03, 0.04 mg/ml) at various concentrations for48 h. The results revealed that both EE and EO induced cell cycle arrest at G1-phase. Cells treated with EE and EO for 48h showed increasing the percentage of cells in G1-phase and decreasing the percentage of cells in S-phase with increasing concentration compared with untreated cells (control). Annexin V-FITC/PI assay confirmed that EO and EE were able to induce apoptosis. Cells treated with EOat (0.04 mg/ml) for 48h resulted in apoptotic cells at 96.68%, and necrotic cells at 0.12%, compared with untreated cells. On the other hand, Cells treated with EE at (0.1 mg/ml) for 48h resulted in apoptotic cells at 94.43%, and necrotic cells at 0.47%, compared with control. Results revealed that EO is better than EE as anticancer; treatment with EO resulted in more apoptotic cells and less necrotic cells, and there were significant differences between them. This confirmed that EO contains specific anticancer compounds as showed by GC-MS analysis. However, more studies should be performed to explore antioxidants present in S.judaica and determine the underlying mechanism of their anti-breast cancer properties.

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