scholarly journals Functional Consequences of Intracellular Proline Levels Manipulation Affecting PRODH/POX–Dependent Pro-Apoptotic Pathways in a Novel in Vitro Cell Culture Model

2017 ◽  
Vol 43 (2) ◽  
pp. 670-684 ◽  
Author(s):  
Ilona Zareba ◽  
Arkadiusz Surazynski ◽  
Marcin Chrusciel ◽  
Wojciech Miltyk ◽  
Milena Doroszko ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Collagen Biosynthesis ◽  
Prolidase Activity ◽  
Proline Level ◽  
Dna Biosynthesis ◽  
Proline Utilization ◽  
Functional Consequences ◽  
Apoptotic Pathways ◽  
Mcf 7

Background/Aims: The effect of impaired intracellular proline availability for proline dehydrogenase/proline oxidase (PRODH/POX)-dependent apoptosis was studied. Methods: We generated a constitutively knocked-down PRODH/POX MCF-7 breast cancer cell line (MCF-7shPRODH/POX) as a model to analyze the functional consequences of impaired intracellular proline levels. We have used inhibitor of proline utilization in collagen biosynthesis, 2-metoxyestradiol (MOE), inhibitor of prolidase that generate proline, rapamycin (Rap) and glycyl-proline (GlyPro), substrate for prolidase. Collagen and DNA biosynthesis were evaluated by radiometric assays. Cell viability was determined using Nucleo-Counter NC-3000. The activity of prolidase was determined by colorimetric assay. Expression of proteins was assessed by Western blot and immunofluorescence bioimaging. Concentration of proline was analyzed by liquid chromatography with mass spectrometry. Results: PRODH/POX knockdown decreased DNA and collagen biosynthesis, whereas increased prolidase activity and intracellular proline level in MCF-7shPRODH/POX cells. All studied compounds decreased cell viability in MCF-7 and MCF-7shPRODH/POX cells. DNA biosynthesis was similarly inhibited by Rap and MOE in both cell lines, but GlyPro inhibited the process only in MCF-7shPRODH/POX and MOE+GlyPro only in MCF-7 cells. All the compounds inhibited collagen biosynthesis, increased prolidase activity and cytoplasmic proline level in MCF-7shPRODH/POX cells and contributed to the induction of pro-survival mode only in MCF-7shPRODH/POX cells. In contrast, all studied compounds upregulated expression of pro-apoptotic protein only in MCF-7 cells. Conclusion: PRODH/POX was confirmed as a driver of apoptosis and proved the eligibility of MCF-7shPRODH/POX cell line as a highly effective model to elucidate the different mechanisms underlying proline utilization or generation in PRODH/POX-dependent pro-apoptotic pathways.

scholarly journals Effect of Hydroalcoholic Extract of Ephedramajor, Memordia charantia, and Resveratrol on Cytotoxicity and Caspase-3 Genes Expression Level in MCF-7 Breast Cancer Cell Line

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Seyed Kazem Sabbagh ◽  
Ehsan Ghodrati ◽  
Alireza Hajibeiki ◽  
Mahta Mazaheri ◽  
Mohammad Reza Sarafraz Ardakani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Medicinal Plants ◽  
Cell Viability ◽  
Cell Line ◽  
Plant Extracts ◽  
Caspase 3 ◽  
Cell Toxicity ◽  
Hydroalcoholic Extract ◽  
Expression Levels ◽  
Mcf 7

Background: To increase the therapeutic effect of drugs to combat diseases, combination therapy with current chemical drugs and new medicines derived from medicinal plants is necessary. Objectives: The present work aimed to investigate the effect of hydroalcoholic extract of two medicinal plants, Ephedra major and Momordi cacharantia (Carla), and resveratrol drug on cell viability and expression levels of caspase-3 gene in MCF-7 cell line. Methods: In this experimental study, the hydroalcoholic extraction of tested plants was done with a Soxhlet extractor. The MTT assay and real-time PCR were used to determine cell toxicity and caspase-3 gene expression levels, respectively. Results: The highest and lowest cytotoxic effects of plant extracts and resveratrol were observed at concentrations of 500 and 150 µg/mL, respectively. The highest level of the caspase-3 gene expression was observed after 72 h of incubation by different concentrations of plant extracts and resveratrol. Conclusions: It can be concluded that both plant extracts could influence cell viability in MCF-7 cells via the increase of cell toxicity and expression of caspase3 gene. Thus, these species could be used in the pharmaceutical industry.

scholarly journals Effect of S-Allyl –L-Cysteine on MCF-7 Cell Line 3-Mercaptopyruvate Sulfurtransferase/Sulfane Sulfur System, Viability and Apoptosis

2020 ◽  
Vol 21 (3) ◽  
pp. 1090 ◽  
Author(s):  
Patrycja Bronowicka-Adamska ◽  
Anna Bentke ◽  
Małgorzata Lasota ◽  
Maria Wróbel
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Breast Adenocarcinoma ◽  
Sulfane Sulfur ◽  
Mercaptopyruvate Sulfurtransferase ◽  
Late Apoptosis ◽  
Sulfur Level ◽  
H2s Production ◽  
Level Reduction ◽  
Mcf 7

The S-Allyl-L-cysteine (SAC) component of aged garlic extract (AGE) is proven to have anticancer, antihepatotoxic, neuroprotective and neurotrophic properties. γ-Cystathionase (CTH), cystathionine β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (MPST) are involved in H2S/sulfane sulfur endogenous formation from L-cysteine. The aim of the study was to determine the effect of SAC on MCF-7 cells survival and apoptosis, which is a widely known approach to reduce the number of cancer cells. An additional goal of this paper was to investigate the effect of SAC on the activity and expression of enzymes involved in H2S production. The experiments were carried out in the human breast adenocarcinoma cell line MCF-7. Changes in the cell viability were determined by MTT assay. Cell survival was determined by flow cytometry (FC). Changes in enzymes expression were analyzed using Western blot. After 24 h and 48 h incubation with 2245 µM SAC, induction of late apoptosis was observed. A decrease in cell viability was observed with increasing SAC concentration and incubation time. SAC had no significant cytotoxic effect on the MCF-7 cells upon all analyzed concentrations. CTH, MPST and CBS expression were confirmed in non-treated MCF-7 cells. Significant decrease in MPST activity at 2245 µM SAC after 24 h and 48 h incubation vs. 1000 µM SAC was associated with decrease in sulfane sulfur levels. The presented results show promising SAC effects regarding the deterioration of the MCF-7 cells’ condition in reducing their viability through the downregulation of MPST expression and sulfate sulfur level reduction.

scholarly journals The Synergistic Combination of Cisplatin and Piperine Induces Apoptosis in MCF-7 Cell Line

2021 ◽  
Author(s):  
Abolfazl Fattah ◽  
Ali Morovati ◽  
Zahra Niknam ◽  
Ladan Mashouri ◽  
Amirhooman Asadi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Viability ◽  
Cancer Cells ◽  
Cell Line ◽  
Cancer Cell ◽  
Breast Cancer Cells ◽  
Caspase 3 ◽  
Assay Method ◽  
Caspase 9 ◽  
Mcf 7

Background: Piperine is a natural compound obtained from the Piper nigrum that exhibits anti-proliferative and anti-cancer activity in cancer cell lines. We analyzed the cytotoxic effect of piperine combined with cisplatin compound in the human MCF-7 breast cancer cell line and the underlying mechanism. Methods: The present in vitro study was performed on MCF-7 cell line in Jahrom University of Medical Sciences between, Jahrom, Iran from 2016 to 2017. Cultured MCF-7 cells were seeded into four groups: a control group (untreated group), a group treated with cisplatin, a group treated with piperine and a group treated with cisplatin and piperine. Cell viability was analyzed using the MTT assay method. Flow c-ytometric analysis was investigated for apoptosis. The mRNA and protein expression of the apoptotic regulators p53, Bcl-2, Bax, caspase 3 and caspase 9 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting analysis. Results: Piperine (20 and 30 µM) in combination with cisplatin (5, 10 and 15 µM) for 24 h synergistically inhibited cell viability of MCF-7 breast cancer cells more than piperine and cisplatin used alone. Synergistic antibreast cancer activities cisplatin (5 µM) and piperine (20 µM) were via inducing apoptosis. Piperine (20 µM) and cisplatin (5 µM) for 24 h induce apoptosis strongly through reduction of Bcl-2 and increase of caspase 3, p53, caspase 9, and Bax. Conclusion: Piperine in combination with cisplatin could trigger p53-mediated apoptosis more effective than cisplatin alone in MCF-7 breast cancer cells, reducing the toxic dose of cisplatin used in cancer chemotherapy.

scholarly journals Cytotoxic and antioxidant properties in vitro of functional beverages based on blackberry (Rubus glaucus Benth) and soursop (Annona muricata L) pulps

2018 ◽  
Vol 8 (11) ◽  
pp. 531 ◽  
Author(s):  
Alexandra Zambrano ◽  
Rosa Raybaudi-Massilia ◽  
Francisco Arvelo ◽  
Felipe Sojo
Keyword(s):  
Antioxidant Activity ◽  
Breast Carcinoma ◽  
Cell Viability ◽  
Cell Line ◽  
Tumor Cells ◽  
Prostate Carcinoma ◽  
Carcinoma Cells ◽  
Cervix Carcinoma ◽  
Tropical Fruit ◽  
Mcf 7

Background:  There are many kinds of tropical fruit available in Venezuela. Two of these fruits are the focus of our study: blackberry (“mora”) and soursop (“guanábana”). These fruits have extraordinary bioactive components. For example, blackberry has antioxidant compounds such as anthocyanins, which are characteristic of the Rosaceae family. Acetogenins present in the Annonaceae family have been shown to possess cytotoxic properties that act against different types of tumor cells. In previous research, we have discovered how lyophilized soursop pulp has an elevated cytotoxic effect with a IC50 of 7.1940±1.06 in human cervix carcinoma cells (HeLa) and 0.8460±1.29 in human prostate carcinoma cells (PC3).Objective: This study focused on the health benefits and properties of the soursop and the blackberry. Our main focus was to determine the antioxidant and cytotoxic properties in a formulated beverage based on blackberry, soursop, and yogurt containing probiotics and prebiotics.Methods: The research includes the study of soursop pulp (SP), blackberry pulp (BP), and two formulations of the functional beverage selected through a sensorial analysis, F2 (BP + SP + Yogurt + Truvía® + Sacarose) and F3 (BP + SP + Yogurt + Truvía® + Sacarose + Sodium tripolyphosphate). Cell viability of prostate carcinoma cells (PC3), breast carcinoma without over-expression of the HER2/c-erb-2 gene (MCF-7), breast carcinoma in which the HER2/c-erb-2 gene is over-expressed (SKBr3) and healthy cells of human connective tissue used as control (Fibroblasts).  The previous indicated samples were assessed using MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide). The antioxidant activity of the functional beverage was also done using a freshly preparation of 1,1-diphenyl-2-picrylhydrazyl (DPPH). Results: The BP demonstrated the highest cytotoxicity for both lines of breast cancer cell lines, MCF-7 and SKBR3. The values of the minimum concentration required to inhibit 50 % of the cell population (IC50) was 0.12 ± 1.10 and 1.81 ± 1.68% v / v respectively, followed by SP in MCF-7 and PC3 with values of 1.40 ± 1.03 and 1.34 ± 1.06 respectively. At the same time, the effectiveness of the formulations used found that 3.60 ± 1.04% v / v of F2 beverage was necessary to achieve 50 % inhibition of cell viability of MCF-7 line. For the formulation F3, it was necessary to use a concentration of 5.21 ± 1.04% v / v for that tumor cell line. However, the F2 and F3 formulations demonstrated IC50 values of 3.69 ± 1.08% v / v and 2.50 ± 1.08% v / v respectively for the PC3 cell line. On the other hand, the antioxidant capacity of BP and SP reached elevated values at 30 minutes of exposure to DPPH, obtaining a rate of 85.28 ± 0.11 and 80.94 ± 0.07 % respectively by using a concentration of 12.5 %, F2 and F3 formulations also reached values of 83.97 ± 0.46 and 85.62 ± 0.11 % at 100 % concentration of both drinks respectively.Conclusion: We discovered that the cytotoxic activity of both formulations prepared as well as the pulps were fairly good, revealing highly effective consequences for the inactivation of breast tumor cells MCF-7 and prostate tumor cells PC3. Moreover, BP and SP demonstrated a high antioxidant activity, with a synergistic effect accomplished by the mixture on F2 and F3.Keywords: Functional beverage, cytotoxic, antioxidant, soursop, blackberry, yogurt.

Determining the Interaction Behavior of Calf Thymus DNA with Anastrozole in the Presence of Histone H1: Spectroscopies and Cell Viability of MCF-7 Cell Line Investigations

2021 ◽  
Author(s):  
Najmeh Zare-Feizabadi ◽  
Zeinab Amiri-Tehranizadeh ◽  
Atena Sharifi-Rad ◽  
Parisa Mokaberi ◽  
Niknaz Nosrati ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Histone H1 ◽  
Calf Thymus Dna ◽  
Interaction Behavior ◽  
Calf Thymus ◽  
Mcf 7

Comparison of the effects of nobiletin and letrozole on the activity and expression of aromatase in the MCF-7 breast cancer cell line

2017 ◽  
Vol 95 (4) ◽  
pp. 468-473 ◽  
Author(s):  
Seyedeh Tayebeh Rahideh ◽  
Mohammad Keramatipour ◽  
Mitra Nourbakhsh ◽  
Fariba Koohdani ◽  
Mostafa Hoseini ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Viability ◽  
Cell Line ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Line ◽  
Cancer Cell Line ◽  
Dependent Manner ◽  
Mtt Assays ◽  
Mcf 7

Nobiletin (NOB) is one of the polymethoxyflavones mainly found in citrus fruits. Aromatase or cytochrome P450 (CYP19) enzyme catalyzes the last and rate-limiting step in estrogen biosynthesis. This study was carried out to investigate the effect of NOB on the activity and expression of aromatase, and to compare this property with letrozole (LET) as aromatase inhibitor in the MCF-7 breast cancer cell line. Cell viability was assessed with 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assays. Aromatase enzyme activity based on the conversion of androgenic substrate testosterone into 17β-estradiol was determined. CYP19 gene expression was measured by quantitative real-time PCR. MTT assays demonstrated that NOB at a concentration of 100 μmol/L decreased cell viability in a time-dependent manner (P < 0.05). NOB significantly inhibited aromatase at the concentration of 0.1 μmol/L (P = 0.013), whereas other concentrations had no effect. Treatment with 10 μmol/L and 1 μmol/L of NOB for 48 h significantly increased (P = 0.001) and decreased (P = 0.02) relative aromatase expression, respectively. The combination of LET and NOB had no effect on aromatase. This study showed for the first time that NOB decreases the activity and expression of aromatase at low concentrations in MCF-7 breast cancer cells.

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