Effects of novel phenolic chalcone derivatives upon MCF-7 Cell viability

Nowadays, although chemotherapy is an established therapy for breast cancer, the molecular mechanisms of chemotherapy resistance in breast cancer remain poorly understood. This study aims to explore the effects of endoplasmic reticulum stress on autophagy, apoptosis, and chemotherapy resistance in human breast cancer cells by regulating PI3K/AKT/mTOR signaling pathway. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to detect the cell viability of six human breast cancer cell lines (MCF-7, ZR-75-30, T47D, MDA-MB-435s, MDA-MB-453, and MDA-MB-231) treated with tunicamycin (5 µM), after which MCF-7 cells were selected for further experiment. Then, MCF-7 cells were divided into the control (without any treatment), tunicamycin (8 µ), BEZ235 (5 µ), and tunicamycin + BEZ235 groups. Cell viability of each group was testified by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Western blotting was applied to determine the expressions of endoplasmic reticulum stress and PI3K/AKT/mTOR pathway–related proteins and autophagy- and apoptosis-related proteins. Monodansylcadaverine and Annexin V–fluorescein isothiocyanate/propidium iodide staining were used for determination of cell autophagy and apoptosis. Furthermore, MCF-7 cells were divided into the control (without any treatment), tunicamycin (5 µM), cisplatin (16 µM), cisplatin (16 µM) + BEZ235 (5 µM), tunicamycin (5 µM) + cisplatin (16 µM), and tunicamycin (5 µM) + cisplatin (16 µM) + BEZ235 groups. Cell viability and apoptosis were also evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Annexin V–fluorescein isothiocyanate/propidium iodide staining. In MCF-7 cells treated with tunicamycin, cell viability decreased significantly, but PEAK, eIF2, and CHOP were upregulated markedly and p-PI3K, p-AKT, and p-MTOR were downregulated in dose- and time-dependent manners. In the tunicamycin + BEZ235 group, the cell viability was lower and the apoptosis rate was higher than those of the control and monotherapy groups. Compared with the cisplatin group, the tunicamycin + cisplatin group showed a relatively higher growth inhibition rate; the growth inhibition rate substantially increased in the tunicamycin + cisplatin + BEZ235 group than the tunicamycin + cisplatin group. The apoptosis rate was highest in tunicamycin + cisplatin + BEZ235 group, followed by tunicamycin + cisplatin group and then cisplatin group. Our study provide evidence that endoplasmic reticulum stress activated by tunicamycin could promote breast cancer cell autophagy and apoptosis and enhance chemosensitivity of MCF-7 cells by inhibiting the PI3K/AKT/mTOR signaling pathway.

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Potential Metabolite Nymphayol Isolated from Water Lily (Nymphaea stellata) Flower Inhibits MCF-7 Human Breast Cancer Cell Growth via Upregulation of Cdkn2a, pRb2, p53 and Downregulation of PCNA mRNA Expressions

Metabolites ◽

10.3390/metabo10070280 ◽

2020 ◽

Vol 10 (7) ◽

pp. 280

Author(s):

Laila Naif Al-Harbi ◽

Pandurangan Subash-Babu ◽

Manal Abdulaziz Binobead ◽

Maha Hussain Alhussain ◽

Sahar Abdulaziz AlSedairy ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Suppressor ◽

Cell Viability ◽

Cancer Cell ◽

Breast Cancer Cell ◽

Kinase Inhibitor ◽

B Cell Lymphoma ◽

Nuclear Antigen ◽

Mrna Levels ◽

Mcf 7

Controlled production of cyclin dependent kinases (CDK) and stabilization of tumor suppressor genes are the most important factors involved in preventing carcinogenesis. The present study aimed to explore the cyclin dependent apoptotic effect of nymphayol on breast cancer MCF-7 cells. In our previous study, we isolated the crystal from a chloroform extract of Nymphaea stellata flower petals and it was confirmed as nymphayol (17-(hexan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol) using x-ray diffraction (XRD), Fourier transform infrared (FTIR), and mass spectroscopy (MS) methods. The cytotoxic effect of nymphayol on MCF-7 cells were analyzed using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The cellular and nuclear damage was determined using propidium iodide (PI) and acridine orange/ethidium bromide (AO/ErBr) staining. Tumor suppressor and apoptosis related mRNA transcript levels were determined using real-time polymerase chain reaction (RT-PCR). Nymphayol potentially inhibits MCF-7 cell viability up to 78%, and the IC50 value was observed as 2.8 µM in 24 h and 1.4 µM in 48 h. Treatment with nymphayol significantly increased reactive oxygen species (ROS) level and the tunnel assay confirmed DNA damage. We found characteristically 76% apoptotic cells and 9% necrotic cells in PI and AO/ErBr staining after 48 h treatment with 2.8 µM of nymphayol. Gene expression analysis confirmed significantly (p ≤ 0.001) increased mRNA levels of cyclin dependent kinase inhibitor 2A (Cdkn2a), retinoblastoma protein 2 (pRb2), p53, nuclear factor erythroid 2-factor 2 (Nrf2), caspase-3, and decreased B-cell lymphoma 2 (Bcl-2), murine double minute 2 (mdm2), and proliferating cell nuclear antigen (PCNA) expression after 48 h. Nymphayol effectively inhibited breast cancer cell viability, and is associated with early expression of Cdkn2a, pRb2, and activation of p53 and caspases.

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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

Gene Cell and Tissue ◽

10.5812/gct.110658 ◽

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.

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Evaluation of Benzamide-chalcone Derivatives as EGFR/CDK2 inhibitor: Synthesis, in-vitro Inhibition, and Molecular Modeling Studies

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520621666210415091359 ◽

2021 ◽

Vol 21 ◽

Author(s):

Akshada Joshi ◽

Heena Bhojwani ◽

Ojas Wagal ◽

Khushboo Begwani ◽

Urmila Joshi ◽

...

Keyword(s):

Molecular Docking ◽

Cell Line ◽

Antiproliferative Activity ◽

Docking Studies ◽

Wild Type ◽

Chalcone Derivatives ◽

Ht 29 ◽

Egfr Kinase ◽

Mcf 7 ◽

Mutant Egfr

Background: EGFR (Epidermal Growth Factor Receptor) and CDK2 (Cyclin Dependent Kinase 2) are important targets in the treatment of many solid tumors and different ligands of these receptors share many common structural features. Objective: The study involved synthesis of benzamide-substituted chalcones and determination of their antiproliferative activity as well as preliminary evaluation of EGFR and CDK2 inhibitory potential using both receptor binding and computational methods. Methods: We synthesized 13 benzamide-substituted chalcone derivatives and tested their antiproliferative activity against MCF-7, HT-29 and U373MG cell-lines using Sulforhodamine B Assay. Four compounds were examined for activity against EGFR and CDK2 kinase. The compounds were docked into both EGFR and CDK2 using Glide software. The stability of the interactions for most active compound was evaluated by Molecular Dynamics Simulation using Desmond software. Molecular Docking studies on mutant EGFR (T790M, T790M/L858R, and T790M/C797S) were also carried out. Results: From the SRB assay, we concluded that compounds 1g, and 1k were effective in inhibiting the growth of MCF-7 cell line whereas the other compounds were moderately active. Most compounds were either moderately active or inactive on U373 MG and HT-29 cell line. Compounds 1g and 1k showed good inhibitory activity against CDK2 kinase while 1d and 1f were moderately active. Compounds 1d, 1f, 1g, and 1k were moderately active against EGFR kinase. Molecular docking reveals involvement of one hydrogen bond with Met793 in binding with EGFR however; it was not stable during simulation and these compounds bind to the receptor mainly via hydrophobic contacts. This fact also points towards a different orientation of the inhibitor within the active site of EGFR kinase. Binding mode analysis for CDK2 inhibition studies indicate that hydrogen bonding interaction with Lys 33 and Leu83 are important for the activity. These interactions were found to be stable throughout the simulation. Considering the results for wild-type EGFR inhibition, the docking studies on mutants were performed and which indicate that the compounds bind to the mutant EGFR but the amino acid residues involved are similar to the wild-type EGFR and therefore, the selectivity seems to be limited. Conclusion: These benzamide-substituted chalcone derivatives will be useful as lead molecules for the further development of newer inhibitors of EGFR and/or CDK2 kinases.

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

International Journal of Molecular Sciences ◽

10.3390/ijms21031090 ◽

2020 ◽

Vol 21 (3) ◽

pp. 1090 ◽

Cited By ~ 1

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.

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Cytotoxic activity of crude saponins from Gaultheria trichophylla against human breast cancer cells MCF-7 and MDA-MB-468

Bangladesh Journal of Pharmacology ◽

10.3329/bjp.v10i2.22992 ◽

2015 ◽

Vol 10 (2) ◽

pp. 443

Author(s):

Fiaz Alam ◽

Qazi Najam us Saqib ◽

Abdul Waheed

Keyword(s):

Breast Cancer ◽

Cell Viability ◽

Crude Extract ◽

Human Breast Cancer ◽

Neutral Red ◽

Breast Cancer Cell Lines ◽

Dependent Manner ◽

Human Breast ◽

Uptake Assay ◽

Mcf 7

This study was conducted to evaluate Gaultheria trichophylla crude extract and respective saponins fraction against human breast cancer cell lines. In MTT assay, cell viability was inhibited by G. trichophylla crude extract (500 µg/mL) and saponins (200 µg/mL) in a dose dependent manner with maximum inhibition of (82% and 85%) and (71% and 42%) against MCF-7 and MDA MB-468, respectively. In neutral red uptake assay, the cell viability was inhibited by crude extract and saponins (100 µg/mL) in a similar manner with maximum inhibitions of (96% and 93%) and (87% and 61%) against MCF-7 and MDA MB-468, respectively, with respect to 91% and 93% inhibition by actinomycin-D (4 µM). The DAPI (4',6-diamidino-2-phenylindole) (10 µg/mL) staining of MCF-7 cells treated with crude saponins showed shrunken nuclei with apparent nuclear fragmentation indicating apoptosis and in contrast, MDA MB-468 showed necrosis mode of cell death. The study exhibited that the G. trichophylla provides new evidences to further explore this plant for the novel targets in anticancer drug development.

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Manganese-Doped Cerium Oxide Nanocomposite Induced Photodynamic Therapy in MCF-7 Cancer Cells and Antibacterial Activity

BioMed Research International ◽

10.1155/2019/7156828 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 9

Author(s):

M. Atif ◽

Seemab Iqbal ◽

M. Fakhar-E-Alam ◽

M. Ismail ◽

Qaisar Mansoor ◽

...

Keyword(s):

Antibacterial Activity ◽

Photodynamic Therapy ◽

Cell Viability ◽

Cerium Oxide ◽

Bacterial Infections ◽

Reactive Oxygen Species Generation ◽

Doped Ceria ◽

Bacterial Strains ◽

Chemical Route ◽

Mcf 7

In this experimental approach, we explored the structures, morphologies, phototoxicities, and antibacterial activities of undoped and Mn-doped ceria nanocomposite materials, MnxCe1−xO2. The MnxCe1−xO2 nanocomposites were synthesized by employing a soft chemical route. Our prime focus was on the influence of different factors, both physical and chemical, i.e., the concentration of manganese in the product, size of the nanocomposite, drug dose, and incubation time, on the bacterial strains. Different bacterial strains were selected as experimental biological models of the antibacterial activity of the manganese-doped cerium oxide nanocomposite. In addition to the photodynamic response, the adenocarcinoma cell line (MCF-7) was also studied. Based on cell viability losses and bacterial inhibition analyses, the precise mechanisms of apoptosis or necrosis of 5-ALA/PpIX-exposed MCF-7 cells under 630 nm red lights and under dark conditions were elucidated. It was observed that the undoped nanocomposites had lower cytotoxicities and inhibitions compared with those of the doped nanocomposites towards pathogens. The antibacterial activity and effectiveness for photodynamic therapy were enhanced in the presence of the manganese-doped ceria nanocomposite, which could be attributed to the correlation of the maximum reactive oxygen species generation for targeted toxicity and maximum antioxidant property in bacteria growth inhibition. The optimized cell viability dose and doping concentration will be beneficial for treating cancer and bacterial infections in the future.

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Comparison on the Cancer Specific Cytotoxicity of Three Gingers (Zingiber officinale Rosc) Leaves Varieties from Indonesia

International Journal of Pharmacognosy and Phytochemical Research ◽

10.25258/ijpapr.v9i1.8053 ◽

2017 ◽

Vol 9 (1) ◽

Author(s):

Susanti S. ◽

Kumoro C. A. ◽

Santoso I. S. ◽

Murwanti R. ◽

Suzery M. ◽

...

Keyword(s):

Cell Viability ◽

Cancer Cell ◽

Detrimental Effect ◽

Zingiber Officinale ◽

Assay Method ◽

Human Breast ◽

Normal Cells ◽

Specific Cytotoxicity ◽

Reduce Cell Viability ◽

Mcf 7

This study was performed to get more insight the cancer specific cytotoxicity of ginger leaf (GL). Three GL varieties (Gajah, Emprit, and Red) were extracted and fractionated. Each etyl acetate fraction in concentration of 200 µg/ml was tested about specific cytotoxicity toward cancer dan normal cells. Cancer cell lines used in this study were human colorectal (HCT116) and human breast (MCF-7 and T47D) cancer while normal cell line was human fibroblast (KMST-6). Based MTS assay method, the results showed Gajah and Emprit GL more significantly reduce cell viability of HCT116 and T47D than Red GL although there was no difference on the efficacy of both varieties. All varieties of GL also significantly reduce cell viability of MCF-7 compare to PBS control. However, there were not significant differences between those GL varieties on their effectiveness against MCF-7. In contrast, there were no effects on the KMST-6 due to all GL varieties treatment compare with PBS control. All data suggested that GL treatment only inhibited in the cancer cells without detrimental effect in the normal cells. Effectiveness of GL against cancer cell varies depend on the varieties. Gajah and Emprit GL are better varieties possess the cancer specific cytotoxicity that merits to be developed as promising chemo preventive agent in the future.

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Effects on Cell Viability and on Apoptosis in Tumoral (MCF-7) and in Normal (MCF10A) Epithelial Breast Cells after Human Chorionic Gonadotropin and Derivated-Angiotensin Peptides Treatments

Journal of Cancer Therapy ◽

10.4236/jct.2013.47a010 ◽

2013 ◽

Vol 04 (07) ◽

pp. 65-69 ◽

Cited By ~ 3

Author(s):

Silvana Aparecida Alves Corrêa de Noronha ◽

Werica Bernardo ◽

Alexandre Jesus Barros ◽

Clovis Ryuichi Nakaie ◽

Suma Imura Shimuta ◽

...

Keyword(s):

Cell Viability ◽

Chorionic Gonadotropin ◽

Human Chorionic Gonadotropin ◽

Angiotensin Peptides ◽

Breast Cells ◽

Mcf 7

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Modeling of antiproliferative effects of Salvia officinalis L. essential oil optimized using Box–Behnken design

10.21203/rs.3.rs-44076/v1 ◽

2020 ◽

Author(s):

Imen Kallel ◽

Ahmed Bayoudh ◽

Bochra Gargouri ◽

Lamia Khannous ◽

Asma Elaguel ◽

...

Keyword(s):

Essential Oil ◽

Cell Viability ◽

Cell Lines ◽

Human Cancer ◽

Salvia Officinalis ◽

Dependent Manner ◽

Box Behnken Design ◽

Cytotoxicity Activity ◽

Mcf 7

Abstract Background Salvia officinalis L. essential oil (SoEO) was mostly traditionally used to medicate various diseases as cancer. Then, the present work aims were: (1) to model the cytotoxicity effects of Salvia officinalis L. essential oil (SoEO) related to the human cancer cell lines kind (MCF-7 and HeLa) ; (2) to optimize the hydro-distillation extraction conditions of SoEO; and, (3) to determine the in vitro scavenging capacity of the free radicals DPPH•, NO•, ABTS+, and the ability to reduce Fe3+. Methods The cytotoxicity and anti-proliferative abilities were evaluated by measuring cell viability and then modeled. Two human cell lines: MCF-7 and HeLa were used. The optimization of SoEO extraction by hydro-distillation was carried out with Response Surface Methodology (RSM) using the Box–Behnken design Results The cytotoxicity activity against both tumor cell lines MCF-7 and HeLa was considerably important with IC50 = 3.125 and 8.920 µg/mL, respectively. All treated cell lines showed a significant reducing in cell viability in response to the increasing oil concentration. The relative behaviors of both cell lines under SoEO treatment were modeled. The obtained optimal extraction yield was Y = 1.85 g/100 g d.b. The main identified fractions were camphene (23.7%), α-thujone (19.62%), 1,8-cineole (10.6%), viridiflorol (5.9%), borneol (5.72%); β-thujone (5.4%); caryophyllene (3,83%). Also, SoEO was mostly able to scavenge DPPH• free radical, ABTS+ radical and hydrogen peroxide in an amount dependent manner (IC50 = 0.97, 0.279 and 0.05 mg/mL, respectively). Conclusion The present work provides a preliminary platform for further investigation of the possible mechanism of S. officinalis essential oils and their individual compounds in cytotoxic and antitumor activity.

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