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Molecular and Cellular Biochemistry Jan 2017Calyptranthes tricona is a species (Myrtaceae) native to South Brazil. Plants belonging to this family are folkloric used for analgesia, inflammation, and infectious...
Calyptranthes tricona is a species (Myrtaceae) native to South Brazil. Plants belonging to this family are folkloric used for analgesia, inflammation, and infectious diseases. However, little is known about the toxic potential of C. tricona. The present study aimed to evaluate the antioxidant activity of C. tricona ethanol and hexane leaf extracts, as well as verify their effect on human lymphocytes and MCF-7 cells. The extracts were subjected to preliminary phytochemical screening, antioxidant activity using DPPH and ORAC methods. Genotoxic and mutagenic effects in cultured human lymphocytes were assessed using the comet assay and the micronucleus assay, respectively. In addition, cell viability by MTT assay and fluorometric analysis of mitochondrial potential and caspases-9 activity were performed in order to verify the possible effects of both extracts on HO-induced cell death of MCF-7 cells. Our findings revealed that the phenol content and the antioxidant activity were only present in the ethanol extract. Also, the phytochemical screening presented steroids, triterpenoids, condensed tannins, and flavones as the main compounds. However, both extracts were capable of inducing concentration-dependent DNA damage in human lymphocytes. When treating MCF-7 cells with the extracts, both of them inhibited MCF-7 cell death in response to oxidative stress through a decrease of mitochondrial depolarization and caspases-9 activity. Thus, our results need to be considered in future in vitro and in vivo studies of C. tricona effects. In the meanwhile, we recommend caution in the acute/chronic use of this homemade preparation for medicinal purpose.
Topics: Cell Death; DNA Damage; Female; Humans; Hydrogen Peroxide; Lymphocytes; MCF-7 Cells; Myrtaceae; Oxidative Stress; Plant Extracts
PubMed: 27704465
DOI: 10.1007/s11010-016-2840-9 -
Journal of the Egyptian National Cancer... Dec 2022Mammosphere formation assay has become a versatile tool to quantify the activity of putative breast cancer stem cells in non-adherent in vitro cultures. However,...
BACKGROUND
Mammosphere formation assay has become a versatile tool to quantify the activity of putative breast cancer stem cells in non-adherent in vitro cultures. However, optimizing the suspension culture system is crucial to establish mammosphere cultures from primary breast tumors.
METHODS
This study aimed at determining the self-renewal and sphere-forming potential of breast cancer stem-like cells derived from human primary invasive ductal carcinoma and normal breast tissue samples, and MCF-7 breast cancer cell line using an optimal suspension culture system. Mammosphere-forming efficiency of the mammospheres generated from the tissue samples and cell line were compared. We evaluated the expression of CD44/CD24/ and CD49f/EpCAM/ phenotypes in the stem-like cells by flow cytometry. CK-18, CK-19, α-SMA, and EpCAM marker expression was assessed using immunohistochemical staining.
RESULTS
Breast epithelial cells isolated from the three samples formed two-dimensional spheroids in suspension cultures. Interestingly, mammospheres formed from patient-derived primary breast tumors were enriched in breast cancer stem-like cells with the phenotype CD44/CD24/ and exhibited a relatively more number of large spheres when compared to the normal breast stem cells. MCF-7-derived SCs were more aggressive and resulted in the formation of a significantly higher number of spheroids. The expression of CK-18/CK-19 and α-SMA/EpCAM proteins was confirmed in breast cancer tissues.
CONCLUSIONS
Thus, the use of primary tumor specimens and breast cancer cell lines as suitable models for elucidating the breast cancer stem cell activity was validated using mammosphere culture system.
Topics: Humans; Female; MCF-7 Cells; Breast; Breast Neoplasms; Neoplastic Stem Cells
PubMed: 36504339
DOI: 10.1186/s43046-022-00152-1 -
European Journal of Gynaecological... 2015To observe the effects of cyclopamine on the biological characteristics of human breast cancer MCF-7 cell line and explore its mechanism.
PURPOSE
To observe the effects of cyclopamine on the biological characteristics of human breast cancer MCF-7 cell line and explore its mechanism.
MATERIALS AND METHODS
After human breast cancer MCF-7 cells were treated with different-concentration cyclopamine for different periods, MTT assay was used to detect the inhibitory effect of cyclopamine on MCF-7 cell proliferation, flow cytometry was used to determine the distribution of MCF-7 cell cycle and the effect of cyclopamine on MCF-7 apoptosis, and Western blot was used to measure the protein levels of cyclins D1 and p21 in MCF-7 cells.
RESULTS
In certain range, MCF-7 cell proliferation was inhibited by cyclopamine in a dose- and time-dependent manner, and the optimal inhibiting concentration was ten µmol/L and the optimal action time at 48 hours. With the time prolongation of cyclopamine action, the cells in G0/G1 phase were significantly increased, but the cells in S phase were significantly decreased (compared with blank control group, allp < 0.05). With the time prolongation of cyclopamine action, apoptosis rate of MCF-7 cells was also significantly increased (compared with blank control group, allp < 0.05). The level of cyclin D1 of MCF-7 cells was decreased, but cyclin p21 was increased (compared with blank control group, all p < 0.05).
CONCLUSION
Cyclopamine inhibits MCF-7 cell proliferation via arresting MCF-7 cell transformation from G1 phase to S phase. This may be associated with the expressions of Hedgehog (Hh) signaling pathway-related cyclins.
Topics: Apoptosis; Breast Neoplasms; Cell Cycle; Cell Proliferation; Cyclin D1; Dose-Response Relationship, Drug; Female; Flow Cytometry; Hedgehog Proteins; Humans; MCF-7 Cells; Veratrum Alkaloids
PubMed: 26390705
DOI: No ID Found -
Journal of Toxicology and Environmental... 2015A recent in vitro study reported that the photoinitiator 2-isopropylthioxanthone (2-ITX) is an endocrine-disrupting compound (EDC). However, it is not clear whether...
A recent in vitro study reported that the photoinitiator 2-isopropylthioxanthone (2-ITX) is an endocrine-disrupting compound (EDC). However, it is not clear whether other photoinitiators such as 1-hydroxycyclohexyl phenyl ketone (1-HCHPK) and 2-methyl-4'-(methylthio)-2-morpholinopropiophenone (MTMP) produce endocrine-disrupting effects. The purpose of this study was thus to assess the association between estrogenic activity and exposure to photoinitiators. For estimation of the proliferative effect of the photoinitiators, the E-screen assay was used. Six photoinitiators, 2,2-dimethoxy-2-phenylacetophenone (2,2-DMPAP), 2-ethylhexyl 4-(dimethylamino)benzoate (2-EHDAB), 1-HCHPK, 2-ITX, methyl-2-benzoylbenzoate (MBB), and MTMP, significantly increased number of MCF-7 cells, an estrogen-sensitive human breast cancer cell line. In addition, pretreatment with estrogen receptor (ER) antagonists such as clomiphene, tamoxifen, or fulvestrant, significantly reversed the proliferative effect of each photoinitiator. Data demonstrated that the six photoinitiators produced endocrine-disrupting effects and that these photoinitiators interacted with ER as agonists. Evidence indicates that the six photoinitiators demonstrated estrogenic activity via ER as agonists.
Topics: Endocrine Disruptors; Female; Humans; MCF-7 Cells
PubMed: 26692070
DOI: 10.1080/15287394.2015.1094431 -
Pharmaceutical Development and... Dec 2023This research aimed to evaluate the effect of variables on exemestane-loaded bovine serum albumin nanoparticles (EXE-BSA NPs) to improve anti-breast cancer activity....
This research aimed to evaluate the effect of variables on exemestane-loaded bovine serum albumin nanoparticles (EXE-BSA NPs) to improve anti-breast cancer activity. EXE-BSA NPs were optimized using 3 factorial design, wherein the concentration of BSA () and sonication time () were independent variables and particle size () and %w/w entrapment efficiency () were dependent variables. The statistical optimization revealed a significant effect of BSA concentration on both variables, whereas sonication time affected only particle size. The optimized EXE-BSA NPs were spherical with 124.1 ± 2.62 nm particle size, 83.95 ± 1.06% w/w drug entrapment, and exhibited a biphasic release of 100% (w/w) drug over 72 h. The optimized formulation induced cytotoxicity in MCF-7 cell lines with an IC50 value of 21.46 µg/mL by MTT assay, almost half the free drug (54.87 µg/mL). Thus, statistically optimized EXE-BSA NPs were effective in MCF-7 cell lines and can be explored to treat estrogen receptor-positive breast cancer.
Topics: Humans; Female; MCF-7 Cells; Drug Carriers; Serum Albumin, Bovine; Breast Neoplasms; Nanoparticles; Particle Size
PubMed: 37987762
DOI: 10.1080/10837450.2023.2285925 -
Talanta May 2018Bisphenol A (BPA) is a widely used additive in the plastic industry and has been reported to have genotoxicity. A hypothesis that BPA may enhance breast cancer risk...
Bisphenol A (BPA) is a widely used additive in the plastic industry and has been reported to have genotoxicity. A hypothesis that BPA may enhance breast cancer risk through the formation of its metabolic intermediate or DNA adduct has been proposed. In this study, breast cancer cell MCF-7 was cultured and the cellular DNA was extracted from the cells. The adducts of bisphenol A 3,4-quinone (BPAQ) with 2'-deoxyguanosine (dG), calf thymus DNA and MCF-7 cell DNA were investigated. DNA adducts were characterized by using electrospray ionization Orbitrap high-resolution mass spectrometry and tandem mass spectrometry. The BPA-DNA adducts of BPAQ with dG, calf thymus and MCF-7 cell DNA were identified as 3-hydroxy-bisphenol A-N7-guanine (3-OH-BPA-N7Gua). The MS/MS fragmentation pathway of 3-OH-BPA-N7Gua was proposed based on obtained accurate mass data. BPA quinone metabolites can react with MCF-7 cell DNA in vitro. The findings provide evidence that BPA might covalently bind to DNA in MCF-7 cells mediated by quinone metabolites, which may increase our understanding of health risk associated with BPA exposure.
Topics: Animals; Benzhydryl Compounds; Benzoquinones; Cattle; DNA; DNA Adducts; DNA, Neoplasm; Deoxyguanosine; Endocrine Disruptors; Humans; MCF-7 Cells; Phenols; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry
PubMed: 29501196
DOI: 10.1016/j.talanta.2018.02.037 -
Chemical Communications (Cambridge,... Feb 2023In this work, we constructed a novel membrane fusion strategy for extracellular vesicles (EVs) and red blood cell membrane vesicles (RVs). A nanoscale space is formed,...
In this work, we constructed a novel membrane fusion strategy for extracellular vesicles (EVs) and red blood cell membrane vesicles (RVs). A nanoscale space is formed, which can improve the efficiency of the probe reaction with miRNA-21, which allows the fluorescence detection of miRNA-21 in EVs.
Topics: Humans; MCF-7 Cells; Extracellular Vesicles; Erythrocyte Membrane; MicroRNAs
PubMed: 36723001
DOI: 10.1039/d2cc05954a -
Environmental Science and Pollution... Jul 2022The utilization of novel compounds as cancer treatments offers enormous potential in this field. The advantages of nanomedicine-based therapy include efficient cellular...
The utilization of novel compounds as cancer treatments offers enormous potential in this field. The advantages of nanomedicine-based therapy include efficient cellular uptake and selective cell targeting. In this study, we employ selenium nanoparticles' green-synthesized by apigenin (SeNPs-apigenin) to treat breast cancer. We used various assays to show that SeNPs-apigenin can reduce MCF-7 cell viability and trigger apoptosis in vitro. Flow cytometry and PCR methods were used to detect apoptosis, while cell migration and invasion methods were used to quantify the possible effect of SeNPs-apigenin therapy on cell migration and invasion. According to cytotoxicity testing, the SeNPs-apigenin treatment can successfully limit MCF-7 cell proliferation and viability in a concentration-dependent manner. Flow cytometric and PCR analyses revealed that SeNPs-apigenin treatment induced apoptosis in MCF-7 cells, demonstrating that SeNPs-apigenin treatment could directly target Bcl-2, Bax, and caspase-3 and result in the discharge of cytochrome C from mitochondria into the cytosol, accompanied by the initiation of cell death, leading to permanent DNA damage and killing of MCF-7 cells. Furthermore, treatment with SeNPs-apigenin increased reactive oxygen species production and oxidative stress in MCF-7 cells. Our findings indicate that SeNPs-apigenin has cytotoxic potential in the treatment of breast cancer.
Topics: Apigenin; Apoptosis; Breast Neoplasms; Female; Humans; MCF-7 Cells; Nanoparticles; Selenium
PubMed: 35182347
DOI: 10.1007/s11356-022-19166-2 -
International Journal of Molecular... Jun 2023Resistance to the chemotherapeutic agents in the clinical management of cancer remains a significant challenge, and the mechanical environment of cancer cells is one of...
Resistance to the chemotherapeutic agents in the clinical management of cancer remains a significant challenge, and the mechanical environment of cancer cells is one of the major determinants of this. Stiffening of the environment is usually associated with increased chemoresistance of cancer cells, although this process depends on the type of cancer. Breast cancer is the most frequently diagnosed cancer, and more than half a million people die from it each year worldwide. In this study, we used the most frequent (70% of diagnosed cases) breast cancer phenotype, representing the MCF-7 cell line, to investigate the influence of surface stiffness on its sensitivity to one of the most commonly used anticancer drugs-doxorubicin. We showed that the mechanical environment affected MCF-7 proliferation, adhesion, and the expression and activation of mitogen-activated protein kinases (MAPKs). Furthermore, the role of MAPKs in response to doxorubicin was dependent on surface stiffness; nevertheless, surface stiffness did not affect MCF-7 resistance to doxorubicin.
Topics: Humans; Female; MCF-7 Cells; Drug Resistance, Neoplasm; Doxorubicin; Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Gene Expression Regulation, Neoplastic
PubMed: 37373337
DOI: 10.3390/ijms241210192 -
Scientific Reports Jul 2016Cold atmospheric plasma (CAP) has been proposed as a useful cancer treatment option after showing higher induction of cell death in cancer cells than in normal cells....
Cold atmospheric plasma (CAP) has been proposed as a useful cancer treatment option after showing higher induction of cell death in cancer cells than in normal cells. Although a few studies have contributed to elucidating the molecular mechanism by which CAP differentially inhibits cancer cell proliferation, no results are yet to be reported related to microRNA (miR). In this study, miR-19a-3p (miR-19a) was identified as a mediator of the cell proliferation-inhibitory effect of CAP in the MCF-7 breast cancer cell. CAP treatment of MCF-7 induced hypermethylation at the promoter CpG sites and downregulation of miR-19a, which was known as an oncomiR. The overexpression of miR-19a in MCF-7 increased cell proliferation, and CAP deteriorated the effect. The target genes of miR-19a, such as ABCA1 and PTEN, that had been suppressed by miR recovered their expression through CAP treatment. In addition, an inhibitor of reactive oxygen species that is produced by CAP suppressed the effect of CAP on cell proliferation. Taken together, the present study, to the best of authors' knowledge, is the first to identify the involvement of a miR, which is dysregulated by the CAP and results in the anti-proliferation effect of CAP on cancer cells.
Topics: Antineoplastic Agents; Cell Proliferation; Epigenesis, Genetic; Humans; MCF-7 Cells; MicroRNAs; Plasma Gases
PubMed: 27445062
DOI: 10.1038/srep30005