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Molecules (Basel, Switzerland) Aug 2021Breast cancer is one of the most prevalent cancers worldwide usually treated with Tamoxifen. Tamoxifen resistance development is the most challenging issue in an...
High Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes.
Breast cancer is one of the most prevalent cancers worldwide usually treated with Tamoxifen. Tamoxifen resistance development is the most challenging issue in an initially responsive breast tumor, and mechanisms of resistance are still under investigation. The objective of this study is to develop and validate a selective, sensitive, and simultaneous high performance liquid chromatography-tandem mass spectrometry method to explore the changes in substrates and metabolites in supernatant media of developed Tamoxifen resistance MCF-7 cells. We focus on the determination of lactate, pyruvate, and L-glutamine which enables the tracking of changes in metabolic pathways as a result of the resistance process. Chromatographic separation was achieved within 3.5 min. using a HILIC column (4.6 × 100 mm, 3.5 µm particle size) and mobile phase of 0.05 M acetic acid-ammonium acetate buffer solution pH 3.0: Acetonitrile (40:60 /). The linear range was 0.11-2.25, 0.012-0.227, and 0.02-0.20 mM for lactate, pyruvate, and L-glutamine, respectively. Within- and between-run accuracy was in the range 98.94-105.50% with precision (CV, %) of ≤0.86%. The results revealed a significant increase in both lactate and pyruvate production after acquiring the resistant. An increase in L-glutamine levels was also observed and could be attributed to its over production or decline in its consumption. Therefore, further tracking of genes responsible of lactate, pyruvate, and glutamine metabolic pathways should be performed in parallel to provide in-depth explanation of resistance mechanism.
Topics: Calibration; Cell Count; Cell Shape; Chromatography, High Pressure Liquid; Drug Resistance, Neoplasm; Glutamine; Humans; Lactic Acid; MCF-7 Cells; Pyruvic Acid; Reproducibility of Results; Tamoxifen; Tandem Mass Spectrometry
PubMed: 34443413
DOI: 10.3390/molecules26164824 -
Cell Death & Disease Aug 2023Ferroptosis, a form of programed cell death, can be promoted by inhibitors of the xCT transporter (erastin) or GPX4 (RSL3). We found that GPX4, but not the xCT...
Ferroptosis, a form of programed cell death, can be promoted by inhibitors of the xCT transporter (erastin) or GPX4 (RSL3). We found that GPX4, but not the xCT transporter, is selectively elevated in luminal breast cancer. Consistent with this observation, the majority of luminal breast cancer cell lines are exquisitely sensitive to RSL3 with limited sensitivity to erastin. In RSL3-resistant, but not sensitive, luminal breast cancer cell lines, RSL3 induces HER2 pathway activation. Irreversible HER2 inhibitors including neratinib reversed RSL3 resistance in constitutively RSL3-resistant cell lines. Combination treatment with RSL3 and neratinib increases ferroptosis through mitochondrial iron-dependent reactive oxygen species production and lipid peroxidation. RSL3 also activated replication stress and concomitant S phase and G2/M blockade leading to sensitivity to targeting the DNA damage checkpoint. Together, our data support the exploration of RSL3 combined with irreversible HER2 inhibitors in clinical trials.
Topics: Humans; Ferroptosis; Apoptosis; Lipid Peroxidation; Iron; MCF-7 Cells; Membrane Transport Proteins; Neoplasms
PubMed: 37596261
DOI: 10.1038/s41419-023-06042-1 -
BMC Complementary Medicine and Therapies Jan 2020Nigella sativa (NS), a member of family Ranunculaceae is commonly known as black seed or kalonji. It has been well studied for its therapeutic role in various diseases,...
BACKGROUND
Nigella sativa (NS), a member of family Ranunculaceae is commonly known as black seed or kalonji. It has been well studied for its therapeutic role in various diseases, particularly cancer. Literature is full of bioactive compounds from NS seed. However, fewer studies have been reported on the pharmacological activity of proteins. The current study was designed to evaluate the anticancer property of NS seed proteins on the MCF-7 cell line.
METHODS
NS seed extract was prepared in phosphate-buffered saline (PBS), and proteins were precipitated using 80% ammonium sulfate. The crude seed proteins were partially purified using gel filtration chromatography, and peaks were resolved by SDS-PAGE. MTT assay was used to screen the crude proteins and peaks for their cytotoxic effects on MCF-7 cell line. Active Peaks (P1 and P4) were further studied for their role in modulating the expression of genes associated with apoptosis by real-time reverse transcription PCR. For protein identification, proteins were digested, separated, and analyzed with LC-MS/MS. Data analysis was performed using online Mascot, ExPASy ProtParam, and UniProt Knowledgebase (UniProtKB) gene ontology (GO) bioinformatics tools.
RESULTS
Gel filtration chromatography separated seed proteins into seven peaks, and SDS-PAGE profile revealed the presence of multiple protein bands. Among all test samples, P1 and P4 depicted potent dose-dependent inhibitory effect on MCF-7 cells exhibiting IC values of 14.25 ± 0.84 and 8.05 ± 0.22 μg/ml, respectively. Gene expression analysis demonstrated apoptosis as a possible cell killing mechanism. A total of 11 and 24 proteins were identified in P1 and P4, respectively. The majority of the proteins identified are located in the cytosol, associate with biological metabolic processes, and their molecular functions are binding and catalysis. Hydropathicity values were mostly in the hydrophilic range.
CONCLUSION
Our findings suggest NS seed proteins as a potential therapeutic agent for cancer. To our knowledge, it is the first study to report the anticancer property of NS seed proteins.
Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cell Proliferation; Chromatography, Gel; Humans; MCF-7 Cells; Mass Spectrometry; Nigella sativa; Pakistan; Plant Extracts; Plant Proteins; Seeds
PubMed: 32020890
DOI: 10.1186/s12906-019-2804-1 -
Scientific Reports Feb 2023Our work group designed and synthesized a promissory compound N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA). The HO-AAVPA is a HDAC1 inhibitor and antiproliferative...
Our work group designed and synthesized a promissory compound N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA). The HO-AAVPA is a HDAC1 inhibitor and antiproliferative in cancer cell lines. However, HO-AAVPA is poor water solubility and enzymatically metabolized. In this work, the fourth-generation poly(amidoamine) dendrimer (PAMAM-G4) was used as a drug deliver carrier of HO-AAVPA. Moreover, HO-AAVPA and HO-AAVPA-PAMAM complex were submitted to forced degradation studies (heat, acid, base, oxidation and sunlight). Also, the HO-AAVPA-PAMAM-G4 complex was assayed as antiproliferative in a breast cancer cell line (MCF-7). The HO-AAVPA-PAMAM-G4 complex was obtained by docking and experimentally using three pH conditions: acid (pH = 3.0), neutral (pH = 7.0) and basic (pH = 9.0) showing that PAMAM-G4 captureand protect the HO-AAVPA from forced degradation, it is due to sunlight yielded a by-product from HO-AAVPA. In addition, the PAMAM-G4 favored the HO-AAVPA water solubility under basic and neutral pH conditions with significant difference (F = 259.9, p < 0.001) between the slopes of the three conditions being the basic condition which solubilizes the greatest amount of HO-AAVPA. Finally, the HO-AAVPA-PAMAM-G4 complex showed better antiproliferative effects on MCF-7 (IC = 75.3 μM) than HO-AAVPA (IC = 192 μM). These results evidence that PAMAM-G4 complex improve the biological effects of HO-AAVPA.
Topics: Humans; Dendrimers; MCF-7 Cells; Water
PubMed: 36854957
DOI: 10.1038/s41598-023-30144-7 -
Journal of Applied Toxicology : JAT Jul 2017Quassinoids often exhibit antioxidant and antiproliferative activity. Emerging evidence suggests that these natural metabolites also display chemopreventive actions. In...
Quassinoids often exhibit antioxidant and antiproliferative activity. Emerging evidence suggests that these natural metabolites also display chemopreventive actions. In this study, we investigated the potential for the quassinoid glaucarubulone glucoside (Gg), isolated from the endemic Jamaican plant Castela macrophylla (Simaroubaceae), to display potent cytotoxicity and inhibit human cytochrome P450s (CYPs), particularly CYP1A enzymes, known to convert polyaromatic hydrocarbons into carcinogenic metabolites. Gg reduced the viability of MCF-7 breast adenocarcinoma cells (IC = 121 nm) to a greater extent than standard of care anticancer agents 5-fluorouracil, tamoxifen (IC >10 μm) and the tamoxifen metabolite 4-hydroxytamoxifen (IC = 2.6 μm), yet was not cytotoxic to non-tumorigenic MCF-10A breast epithelial cells. Additionally, Gg induced MCF-7 breast cancer cell death. Gg blocked increases in reactive oxygen species in MCF-10A cells mediated by the polyaromatic hydrocarbon benzo[a]pyrene (B[a]P) metabolite B[a]P 1,6-quinone, yet downregulated the expression of genes that promote antioxidant activity in MCF-7 cells. This implies that Gg exhibits antioxidant and cytoprotective actions in non-tumorigenic breast epithelial cells and pro-oxidant, cytotoxic actions in breast cancer cells. Furthermore, Gg inhibited the activities of human CYP1A according to non-competitive kinetics and attenuated the ability of B[a]P to induce CYP1A gene expression in MCF-7 cells. These data indicate that Gg selectively suppresses MCF-7 breast cancer cell growth without impacting non-tumorigenic breast epithelial cells and blocks B[a]P-mediated CYP1A induction. Taken together, our data provide a rationale for further investigations of Gg and similar plant isolates as potential agents to treat and prevent breast cancer. Copyright © 2017 John Wiley & Sons, Ltd.
Topics: Antineoplastic Agents, Phytogenic; Antioxidants; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cytochrome P-450 Enzyme System; Cytotoxins; Female; Gene Expression; Glaucarubin; Humans; Jamaica; MCF-7 Cells; Plant Extracts; Quassins; Simaroubaceae
PubMed: 28138972
DOI: 10.1002/jat.3436 -
Asian Pacific Journal of Cancer... Dec 2020The purpose of this research was to evaluate the cytotoxic, cell cycle arrest, and apoptotic induction activities of the fruit of S. nigrum L. ethanolic-70% extract...
OBJECTIVE
The purpose of this research was to evaluate the cytotoxic, cell cycle arrest, and apoptotic induction activities of the fruit of S. nigrum L. ethanolic-70% extract against MCF-7 human breast cancer cell.
METHODS
S. nigrum L. ripe fruit was blended and macerated with ethanol 70% and the filtrate was evaporated. The semisolid extract was then analyzed phytochemically. Cytotoxic analysis was performed using MCF-7 cancer and Vero normal cell by MTT method and followed by apoptotic and cell cycle arrest analysis using flow cytometry.
RESULTS
The phytochemical analysis resulted that extract contained total phenolic and flavonoid compounds with the level of 1.545±0.080% and 0.212±0.002%, respectively. Glycitin was the highest level of isoflavone compound, namely, 375.0844 mg/100 g extract. The cytotoxic evaluation revealed that the extract exhibited a selectively toxic effect between cancer and normal cell. The extract inhibited MCF-7 proliferation with IC50 value about 40.77±4.86 μg/mL and conversely toward Vero cell at lower cytotoxic activity with an IC50 value of 298.96±27.28 μg/mL. Evaluation of MCF-7 cell cycles demonstrated that the extract arrested the cell cycle in the S phase and continued to the G2/M phase at the half of the IC50 value. The extract induced apoptotic of MCF-7 cell about 43.31% in which this activity was nearly the same with doxorubicin as a positive control (59.14%). However, solamargine was predicted as the most active anticancer compounds by a molecular docking study so that it was suggested to measure the level of this compound.
CONCLUSION
It can be concluded that the fruit of S. nigrum L. ethanolic-70% extract demonstrated cytotoxic activity toward MCF-7 breast cancer cell and nontoxic on Vero normal cell. Solamargine was predicted as the most active anticancer compound. This extract had an opportunity to be developed as a potential anticancer agent to overcome breast cancer diseases.
Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Breast Neoplasms; Cell Cycle Checkpoints; Cell Proliferation; Ethanol; Female; Humans; MCF-7 Cells; Molecular Docking Simulation; Phytotherapy; Plant Extracts; Solanum nigrum
PubMed: 33369475
DOI: 10.31557/APJCP.2020.21.12.3735 -
International Journal of Molecular... Dec 2022The existence of a tight relationship between inflammation and epigenetics that in primary breast tumor cells can lead to tumor progression and the formation of bone...
The existence of a tight relationship between inflammation and epigenetics that in primary breast tumor cells can lead to tumor progression and the formation of bone metastases was investigated. It was highlighted how the induction of tumor progression and bone metastasis by Interleukin-1 beta, in a non-metastatic breast cancer cell line, MCF-7, was dependent on the de-methylating actions of ten-eleven translocation proteins (TETs). In fact, the inhibition of their activity by the Bobcat339 molecule, an inhibitor of TET enzymes, determined on the one hand, the modulation of the epithelial-mesenchymal transition process, and on the other hand, the reduction in the expression of markers of bone metastasis, indicating that the epigenetic action of TETs is a prerequisite for IL-1β-dependent tumor progression and bone metastasis formation.
Topics: Female; Humans; Bone Neoplasms; Breast Neoplasms; Cell Line, Tumor; Epigenesis, Genetic; Epithelial-Mesenchymal Transition; Inflammatory Breast Neoplasms; Interleukin-1beta; MCF-7 Cells; Dioxygenases; DNA-Binding Proteins; Enzyme Inhibitors
PubMed: 36499741
DOI: 10.3390/ijms232315422 -
In Vivo (Athens, Greece) 2020Human mesenchymal stem cells (hMSC) represent a versatile cell population, able to modulate the tumor microenvironment. Our aim was to recreate an open scene for the in...
BACKGROUND/AIM
Human mesenchymal stem cells (hMSC) represent a versatile cell population, able to modulate the tumor microenvironment. Our aim was to recreate an open scene for the in vivo interaction between hMSC and the MCF-7 breast cancer cells (MCF-7), in order to enlighten the intimate involvement of hMSC in tumor vasculogenesis and angiogenesis.
MATERIALS AND METHODS
hMSC and MCF-7 were seeded onto the chick embryo chorioallantoic membrane (CAM) and incubated for 7 days. Consecutively, the morphology and the immunohistochemical profile of CAM were assessed.
RESULTS
Following this complex interaction, MCF-7 acquired a more aggressive phenotype, hMSC switched to a vascular precursor phenotype, while CAM underwent a major reset to an earlier stage, with hotspots of angiogenesis, vasculogenesis and hematopoiesis.
CONCLUSION
The hallmark of this study was the establishment of a veritable in vivo experimental model of MSC involvement in tumor vasculogenesis and angiogenesis, allowing further analysis in the field.
Topics: Animals; Cell Differentiation; Chick Embryo; Chorioallantoic Membrane; Humans; MCF-7 Cells; Neovascularization, Pathologic; Neovascularization, Physiologic
PubMed: 33144439
DOI: 10.21873/invivo.12170 -
Journal of Bioenergetics and... Dec 2020The anti-proliferative activities of Novel series of 2-(4-fluorophenyl) imidazol-5-ones against MCF-7 breast cancer cell line were explored via in-slico studies which...
The anti-proliferative activities of Novel series of 2-(4-fluorophenyl) imidazol-5-ones against MCF-7 breast cancer cell line were explored via in-slico studies which includes Quantitative structure-activity relationship QSAR, molecular docking studies, designing new compounds, and analyzing the pharmacokinetics properties of the designed compounds. From the QSAR analysis, model number one emerged the best as seen from the arithmetic assessments of (R) = 0.6981, (R) = 0.6433, (Q) = 0.5460 and (R) of 0.5357. Model number one was used in designing new derivative compounds, with higher effectiveness against estrogen positive breast cancer (MCF-7 cell line). The Molecular docking studies between the derivatives and Polo-like kinases (Plk1) receptor proved that the derivatives of 2-(4-fluorophenyl) imidazol-5-ones bind tightly to the receptor, thou ligand 24 and 27 had the highest binding affinities of -8.8 and - 9.1 kcal/mol, which was found to be higher than Doxorubicin with a docking score of -8.0 kcal/mol. These new derivatives of 2-(4-fluorophenyl) imidazol-5-ones shall be excellent inhibitors against (plk1). The pharmacokinetics analysis performed on the new structures revealed that all the structures passed the test and also the Lipinski rule of five, and they could further proceed to pre-clinical tests. They both revealed a revolution in medicine for developing novel anti-breast cancer drugs against MCF-7 cell line.
Topics: Breast Neoplasms; Cell Line, Tumor; Female; Humans; Imidazoles; MCF-7 Cells; Molecular Docking Simulation; Quantitative Structure-Activity Relationship
PubMed: 33247393
DOI: 10.1007/s10863-020-09858-0 -
International Journal of Nanomedicine 2020L. is a very important medicinal and industrial flowering aromatic plant.
INTRODUCTION
L. is a very important medicinal and industrial flowering aromatic plant.
METHODS
The present study deals with L. leaves extract (JOLE) as a reducing and capping agent for the synthesis of silver nanoparticles (AgNPs) by the green pathway. Phenolic profile of the extract was evaluated using HPLC-PDA/MS/MS technique. L. leaves extract silver nanoparticles (JOLE-AgNPs) were characterized by ultraviolet light (UV), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), zeta potential and X-ray diffraction (XRD). JOLE-AgNPs were examined for their cytotoxic activities by neutral red uptake assay (NRU) against bladder (5637) and breast cancer (MCF-7) cell lines.
RESULTS
HPLC-PDA/MS/MS tentatively identified 51 compounds of different chemical classes. UV spectra showed absorption peak at λmax = 363 nm. The biosynthesized AgNPs were predominantly spherical in shape with an average size of 9.22 nm by TEM. The face cubic center (fcc) nature of silver nanoparticles was proved by XRD diffractogram. JOLE-AgNPs exhibited high cytotoxic activity against 5637 and MCF-7 cell lines compared to the cytotoxic activities of JOLE with IC of 13.09 µg/mL and 9.3 µg/mL, respectively.
DISCUSSION
The silver nanoparticles formed by L. showed high cytotoxic activities against MCF-7 and 5637 cell lines and can be introduced as a new alternative cytotoxic medication.
Topics: Antineoplastic Agents; Breast Neoplasms; Chemistry Techniques, Synthetic; Green Chemistry Technology; Humans; Jasminum; MCF-7 Cells; Metal Nanoparticles; Plant Extracts; Plant Leaves; Silver; Urinary Bladder
PubMed: 33304101
DOI: 10.2147/IJN.S269880