Cultured MCF-7 Cells Research Articles

Design, synthesis, and biological evaluation of thiazole bioisosteres of goniofufurone through in vitro antiproliferative activity and in vivo toxicity

The synthesis of several new goniofufurone bioisosteres was achieved in which the phenyl residue was replaced by a thiazole ring. The key steps of the synthesis included the initial condensation of cyanohydrin benzoates with cysteine ethyl ester hydrochloride, followed by the subsequent reaction of resulting C-4′ epimeric thiazolines with DBU, to introduce 5-deoxy functionality and to elaborate the thiazole ring in one step. Synthesized compounds showed potent growth inhibitory effects against selected human tumour cell lines, especially bioisostere 4, which in the culture of MCF-7 cells displayed the highest activity (IC50 = 0.19 nM) of all compounds under evaluation. This molecule exhibited 64474-fold higher antiproliferative activity than lead 2 and was1053-fold more active than the commercial antitumour agent doxorubicin in the culture of MCF-7 cells. The structural features of the tested compounds responsible for their antiproliferative activity have been identified by preliminary SAR analysis. The toxicity of the most active compound 4 was assessed by an in vivo experiment in a zebrafish model (Danio rerio), whereupon it was found non-toxic at any of the tested concentrations up to 125 μM.

Abstract P3-10-03: Enrichment of pro-oncogenic immune miRNAs in exosomes by cyclin D1, promote cancer stem cell expansion

Abstract The cyclin D1 (CCND1) gene, encodes the regulatory subunit of a holoenzyme that phosphorylates and inactivates the Rb protein. Cyclin D1 overexpression occurs in up to 50% of human breast cancers. Cyclin D1, governs the expression (1,3), processing (2), the secretion and the relative proportion of secreted non-coding RNA subtypes (miRNA, rRNA, tRNA, CDBox, scRNA, HAcaBox. scaRNA, piRNA) in human breast cancer and the secretion of piRNA (4). Through base-pairing to the complementary sequence in 3’ untranslated region (3’ UTR) of mRNA, miRNAs participate in diverse biological processes. A subset of miRNA regulate the ageing of innate and adaptive immune cells, which has been associated with breast cancer initiation, progression and therapy resistance. Immuno-miRs, ( miR-21 and miR-93), bind to the 3′-UTR region of mRNAs, and binding to- and or activate Toll-Like Receptor 8 (TLR8) in surrounding cells of the immune system. Exosomes which contain constituents of their cell of origin, including mRNA and miRNA are found in the tumor microenvironment and participate in therapy resistance, cancer metastasis and the metastatic niche. Herein, tet-inducible cyclin D1 shRNA knock down in MCF-7 breast cancer cells reduced MCF-7 cell proliferation and mammosphere formation. Exosomes isolated from wildtype and cyclin D1 knock-down MCF-7 cell culture media, showed that exosomes from cyclin D1 knock down cells decreased proliferation of both wildtype and cyclin D1 knock-down cells. Furthermore, wildtype exosomes partially reversed the cyclin D1 knock-down effects. Cyclin D1 knock-down exosomes decreased mammosphere formation of wildtype MCF-7 cells and wildtype exosomes increased mammosphere formation of cyclin D1 knock-down cells. miRNA profiles within the exosomes, showed that cyclin D1 regulated the abundance of miRNA within exosomes. >85% of the cyclin D1-induced exosome miRNA transcripts were the immune-mIR, miR21. hsa-mir-21 which has documented tumor promoting function, was the most enriched miRNAs in cyclin D1 knock-down exosomes. In contrast, exosome abundance of the transcripts for hsa-mir-16 and hsa-mir-92a, both of which were showed as tumor suppressor in breast cancer cells, were reduced by cyclin D1 shRNA. The cyclin D1-mediated miRNA exosome content may contribute to tumor initiation and progression through heterotypic expansion of cancer stem cells. 1.Yu, Z., etal ., A cyclin D1/microRNA 17/20 regulatory feedback loop in control of breast cancer cell proliferation. J Cell Biol. 2008 Aug 11; 182(3): p. 509-17. 2.Yu., Z,., et. al. Cyclin D1 induction of Dicer governs microRNA processing and expression in breast cancer. Nat Commun 2013;4:2812.doi: 10.1038/ncomms3812. 3.Yu Z, et al. Cyclin D1- Mediated MicroRNA Expression Signature in Breast Cancer Subtype and Outcome. Theranostics, 2018 Mar 11,8:8, 2251-2263. 4.Lü J, et al. Cyclin D1 promotes secretion of pro-oncogenic immuno-miRNAs and piRNAs. Clin Sci (Lond). 2020 Mar 27. pii: CS20191318. doi: 10.1042/CS20191318. [Epub ahead of print]PMID:32219337 Citation Format: Xuanmao Jiao, Chongwen Xu, Lifeng Tian, Zhao Zhang, Anthony W Ashton, Zhiping Li, Richard G Pestell. Enrichment of pro-oncogenic immune miRNAs in exosomes by cyclin D1, promote cancer stem cell expansion [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-10-03.

Culture and in situ H2O2-mediated electrochemical study of cancer cells using three-dimensional scaffold based on graphene foam coated with Fe3O4 nanozyme.

For real-time evaluation of the cell behavior and function under in vivo-like 3D environment, the 3D functionalized scaffolds simultaneously integrate the function of 3D cell culture, and electrochemical sensing is a convincing candidate. Herein, Fe3O4 nanoparticles as the nanozyme (peroxide oxidase mimics) were modified on graphene foam scaffold to construct a 3D integrated platform. The platform displayed a wide linear range of 100nM to 20μM and a high sensitivity of 53.2 nA μM-1 toward detection of hydrogen peroxide (H2O2) under the working potential of + 0.6V (vs. Ag/AgCl). The obtained 3D scaffold also displayed satisfactory selectivity toward the possible interferents that appeared in the cell culture environment. Furthermore, the cells still maintained high cell viability (almost 100%) after their growth and proliferation on the scaffold for 7days. With the superior performance on cell culture and electrochemical monitoring, the functions on the 3D culture of MCF-7 or HeLa cells and in situ monitoring of cell-released H2O2 was easily achieved on this 3D platform, which show its great application prospects on further cancer-related disease diagnosis or drug screening. A nanozyme-based three-dimensional graphene scaffold was successfully constructed for cell culture and identification of cancer cells through in situ electrochemical monitoring of the cell-released H2O2.

CYTOTOXIC ACTIVITY OF NEW AZOLOAZINES IN THE STUDY OF HUMAN BREAST CANCER MCF-7 133 CELL CULTURE

The work presents the results of study the cytotoxic activity of new azoloazine derivatives to establish the potential its possibility of use as antitumor agents, including for chemotherapy of metastatic breast cancer. The relevance of the work is due to the wide spread of oncological diseases and high cancer mortality, which dictates the need to constantly obtain cell lines and improve cultivation protocols for testing new antitumor drugs and obtaining new information about the chemistry of cancer. The culturing MCF-7 cells and determining cytotoxicity in a methyltetrazolium test are base methods used in this study. Comparative cytotoxicity was carried out by determining the concentration causing 50 % cell death for 11 azolotriazine derivatives taken at concentrations from 0.25 to 10.0 pM/L. Epirubicin, widely used in breast cancer chemotherapy, we used as a comparison drug. As a result, it was found that not all substances have toxic properties, despite the similar chemical structure and mechanism of action on the cell. The least toxic substances 10 and 11, in which, at the maximum studied concentration of 10 |uM/L, cell survival was 86 and 75 %, respectively. The most toxic compounds against MCF-7 cells are substances 4 and 9 derivatives of with cytotoxicity indicators higher, respectively, by 11.0 and 3.1 highly compared to Epirubicin. The obtained data can be used as a basis for the selection of substances 4 and 9 for further study of their properties on cell models and laboratory animals as substances with potential anticancer activity.

Synthesis, characterization and biological activities of complexes CoII, NiII and CuII prepared using mixture of 2-hydroxy-1-naphthaldehyde and N, N′-dimethyl-1, 3-propanediamine

Copper-based compounds are promising entities for target-specific next-generation anticancer therapeutics. In lieu of this, 2-Hydroxy-1-Naphthaldehyde and N, N′-Dimethyl-1,3-Propanediamine scaffold plays an important role, because of their wide variety of potential (NCS) = Thiocynade and coordination modes. Herein, we report copper complexes 3 with a biocompatible molecule, and secondary ligands viz,. copper complexes have been designed, synthesized and adequately characterized using various spectroscopic techniques. In-vitro, DNA and Bovine serum albumin (BSA) binding was also carried out using fluorescence technique Furthermore, copper complexes 1–3 were tested for biological studies, i.e., anticancer as well as In vitro cytotoxicity results were carried out on cultured MCF-7 cell lines responsible for Moreover, and the copper complexes 3 were tested for potential to act cytotoxicity.

Research possibility of using an enzymatic hydrolyzate of the lymphoid tissue of broiler chickens as an immunomodulator

Currently, various adverse factors can lead to disruption of the immune system. In order to restore the functional state of immunity, it is necessary to use immunomodulators. One such immunomodulators is an enzymatic hydrolyzate of the lymphoid tissue of broiler chickens . The aim of this work was to study the effect of an enzymatic hydrolyzate of the lymphoid tissue of broiler chickens on the morphofunctional state of normal and tumor cells. The object of the research was stem cell culture , human dermal fibroblasts and tumor cell cultures : human cervical carcinoma cells HeLa and adenocarcinoma MCF-7 human breast. The cells were incubated in 96-hole tripod in CO 2 incubator at 37 °C. These cultures were supplemented with dietary supplements in concentrations of 0.05%; 0.1%; 0.5%; 1%; 5%; 10%; 20%; 25%; 40% and 50%. After 48 h, the viability of the cells was examined using MTT-analysis. The experiment revealed cytotoxic effects on mesenchymal stem cells and on MCF-7 breast adenocarcinoma cells. An enzymatic hydrolyzate of the lymphoid tissue of broiler chickens did not have a pronounced effect on viability and did not show a cytotoxic effect in relation to human dermal fibroblasts. In the case of HeLa cells, dietary supplements stimulate their proliferative activity. The response from tumor and stem cells requires further study to identify possible action mechanisms of an enzymatic hydrolyzate of the lymphoid tissue of broiler chickens. • Hydrolysate in concentrations 0.05%; 0.1%; 0.5%; 1%; 5%; 10% it does not reduce the viability of mesenchymal cells. • High doses of hydrolysate 25% and 50% have a weak cytotoxic effect on mesenchymal cells. • Hydrolysate in concentrations of 10%, 20% and 40% has a significant dose-dependent cytotoxic effect on MCF-7 cell culture. • Hydrolysate in concentrations of 5%, 10% and 20% stimulates the proliferative activity of HeLa cells. • The reaction of tumor and stem cells requires further study to identify possible mechanisms of action of the hydrolysate.

Targeted combined therapy in 2D and 3D cultured MCF-7 cells using metformin and erlotinib-loaded mesoporous silica magnetic nanoparticles

Aim This research aims to develop potential therapeutic nanostructures (NSs) encapsulating metformin (MET) and erlotinib (ER) for combinational therapy in breast cancer. Methods The ER and MET, both were loaded on mesoporous silica magnetic nanoparticles conjugated with polyethylene glycol and methotrexate to achieve targeted NSs. The developed NSs were characterised using SEM, DLS, and FTIR. Afterward, MTT, Trypan blue, and DNA extraction assays were operated for biological evaluations in the 2D and 3D MCF-7 cells. Results Physicochemical approaches indicated the mean diameter of 69.4 nm ± 9.5 (PDI = 0.64), and neutral charge (2 mv) for the developed NSs. MET and ER-loaded NSs exhibited 62.56% ± 4.41 and 67.73% ± 3.03 drug release amount in pH = 5.4, respectively. MTT assay revealed that ER- and MET-loaded NSs had less metabolic activity (≈ 20%) in comparison with non-targeted NSs. Conclusion Overall, our combined ER and MET-loaded targeted NSs result in a synergistic inhibitory impact on MCF-7 cells.

Jolkinolide B inhibits proliferation or migration and promotes apoptosis of MCF-7 or BT-474 breast cancer cells by downregulating the PI3K-Akt pathway

Ethnopharmacological relevanceThe diterpenoids extracted from Euphorbia kansui S.L. Liou ex S.B.Ho, Euphorbia fischeriana Steud. have good antitumor effects. Jolkinolide B has anti-breast cancer effect, but it is unclear whether it has different therapeutic effects between luminal A subtype and luminal B subtype breast cancer. Aim of the studyThis study investigated the Jolkinolide B has different therapeutic, important targets and pathways effects between luminal A subtype and luminal B subtype breast cancer. Materials and methodsWe used bioinformatics to predict the biological process and molecular mechanism of Jolkinolide B in treating two types of breast cancer. Then, in vitro, cultured MCF-7 cells and BT-474 cells were divided into control group, PI3K inhibitor + control group, Jolkinolide B group and PI3K inhibitor + Jolkinolide B group. The CCK-8 assay, Flow cytometric analysis and Transwell cell migration assay was used to detect the cell proliferation, apoptosis, and migration in each group, respectively. ELISA was used to measure the content of Akt and phosphorylated Akt (p-Akt) in cell lysis buffer. ResultsCompared to luminal A breast cancer, Jolkinolide B had more targets, proliferation, migration processes and KEGG pathways when treating luminal B subtype breast cancer. Jolkinolide B significantly prolonged the survival time of luminal B subtype breast cancer patients. Compared to the control group, the cell proliferation absorbance value (A value) and migration number of the two kinds of breast cancer cells in the Jolkinolide B group were decreased (P < 0.01, n = 6), and the number of apoptotic cells was increased (P < 0.01, n = 6). Compared to the Jolkinolide B group, the A value and migration number of the two types of breast cancer cells were significantly decreased in the PI3K inhibitor + Jolkinolide B group (P < 0.01, n = 6), and the number of apoptotic cells was significantly increased (P < 0.01, n = 6). In addition, compared to MCF-7 cells, the A value and migration number of BT-474 cells stimulated with Jolkinolide B were significantly decreased (P < 0.01, n = 6), and the number of apoptotic cells was significantly increased (P < 0.01, n = 6). Akt and p-Akt protein levels in the two breast cancer cell lines in the Jolkinolide B group were all decreased (P < 0.01, n = 6), especially in BT-474 cells stimulated by Jolkinolide B. ConclusionJolkinolide B regulates the luminal A and luminal B subtypes of breast cancer through PI3K-Akt, EGFR and other pathways. Jolkinolide B has more significant therapeutic effect on luminal B subtype breast cancer. In vitro, experiments verified that Jolkinolide B significantly inhibited the proliferation and migration activity of BT-474 breast cancer cells by downregulating the PI3K-Akt pathway.

Inhibition of STAT3 enhances sensitivity to tamoxifen in tamoxifen-resistant breast cancer cells

BackgroundThe mechanisms of endocrine resistance are complex, and deregulation of several oncogenic signalling pathways has been proposed. We aimed to investigate the role of the EGFR and Src-mediated STAT3 signalling pathway in tamoxifen-resistant breast cancer cells.MethodsThe ER-positive luminal breast cancer cell lines, MCF-7 and T47D, were used. We have established an MCF-7-derived tamoxifen-resistant cell line (TamR) by long-term culture of MCF-7 cells with 4-hydroxytamoxifen. Cell viability was determined using an MTT assay, and protein expression levels were determined using western blot. Cell cycle and annexin V staining were analysed using flow cytometry.ResultsTamR cells showed decreased expression of estrogen receptor and increased expression of EGFR. TamR cells showed an acceleration of the G1 to S phase transition. The protein expression levels of phosphorylated Src, EGFR (Y845), and STAT3 was increased in TamR cells, while phosphorylated Akt was decreased. The expression of p-STAT3 was enhanced according to exposure time of tamoxifen in T47D cells, suggesting that activation of STAT3 can cause tamoxifen resistance in ER-positive breast cancer cells. Both dasatinib (Src inhibitor) and stattic (STAT3 inhibitor) inhibited cell proliferation and induced apoptosis in TamR cells. However, stattic showed a much stronger effect than dasatinib. Knockdown of STAT3 expression by siRNA had no effect on sensitivity to tamoxifen in MCF-7 cells, while that enhanced sensitivity to tamoxifen in TamR cells. There was not a significant synergistic effect of dasatinib and stattic on cell survival. TamR cells have low nuclear p21(Cip1) expression compared to MCF-7 cells and inhibition of STAT3 increased the expression of nuclear p21(Cip1) in TamR cells.ConclusionsThe EGFR and Src-mediated STAT3 signalling pathway is activated in TamR cells, and inhibition of STAT3 may be a potential target in tamoxifen-resistant breast cancer. An increase in nuclear p21(Cip1) may be a key step in STAT3 inhibitor-induced cell death in TamR cells.

Biofunctional supramolecular hydrogels fabricated from a short self-assembling peptide modified with bioactive sequences for the 3D culture of breast cancer MCF-7 cells

Self-assembling peptides are a type of molecule with promise as scaffold materials for cancer cell engineering. We have reported a short self-assembling peptide, (FFiK)2, that had a symmetric structure connected via a urea bond. In this study, we functionalized (FFiK)2 by conjugation with various bioactive sequences for the 3D culture of cancer cells. Four sequences, RGDS and PHSRN derived from fibronectin and AG73 and C16 derived from laminin, were selected as bioactive sequences to promote cell adhesion, proliferation or migration. (FFiK)2, and its derivatives could co-assemble into supramolecular nanofibers displaying bioactive sequences and form hydrogels. MCF-7 cells were encapsulated in functionalized peptide hydrogels without significant cytotoxicity. Encapsulated MCF-7 cells proliferated under 3D culture conditions. MCF-7 cells proliferated with spheroid formation in hydrogels that displayed RGDS or PHSRN sequences, which will be able to be applied to drug screening targeting cancer stem cells. On the other hand, since MCF-7 cells migrated in a 3D hydrogel that displayed AG73, we could construct the metastatic model of breast cancer cells, which is helpful for the elucidation of breast cancer cells and drug screening against cancer cells under metastatic state. Therefore, functionalized (FFiK)2 hydrogels with various bioactive sequences can be used to regulate cancer cell function for tumor engineering and drug screening.

Abstract 2032: Vitamin B12 augments Ca2+ dependent cytotoxicity of diethyldithiocarbamate through excessive ER-vacuolization, oxidative stress and mitochondrial depolarization in MCF-7 cells in culture

Abstract Coordinated organization of intracellular organelles into functional networks is vital for cells. Mitochondrial and endoplasmic reticulum (ER) interaction in regulation of cell Ca2+ homeostasis is crucial for cell death and survival. Function of these intracellular organelles involved in two-way regulation of the status of major cell metabolites including protein, lipid exchange and intracellular Ca2+ homeostasis. Disulfiram and its derivatives recently attracted attention as a potential anticancer treatment (Kona et al., 2011). Combined use of hydroxycobalamin (vitamin B12b) and diethyldithiocarbamate (DDTC) demonstrated synergistic effect (combination index=0.25) and enhanced the cytotoxic effect of DDTC in number of transformed cell lines in culture (Solovieva et al., 2020). In the present work using MCF-7, human breast cancer cells in culture we demonstrate that combined action of DDTC and B12b resulted in induction of UPRER manifested by dilation of ER cisterns, increased level of polyubiquitinated proteins, and increased expression of CHOP. Confocal microscopy revealed disorganization of mitochondrial network and ER and treatment of these cells with B12b/DDTC and accompanied by mitochondrial depolarization (~50% after 6 h of exposure to drugs). Treatments also induced changes in morphology of mitochondrial network, and within 6 h of exposure, mitochondria demonstrated large amplitude swelling, characteristic feature of mitochondrial permeability transition pore opening. Drug exposure induced mitochondrial oxidative stress and increase in superoxide anion level (monitored with MitoSOX) and increased content of mitochondrial Ca2+ (xRhod-1) which almost doubled following treatment with B12b/DDTC. Short-time exposure of MCF-7 cells to mixture of drugs in Ca2+-free incubation media (2.5 mM EGTA) resulted in generation of significantly smaller size of vacuoles as compared with control. Similarly, pre-loading of cells with intracellular Ca2+ chelator (BAPTA, 5 µM, 30 min, 37oC) or inhibition of IP3 receptor(s) with 2-APB, delayed the drug-induced vacuolization. Taken together, our data indicate that the mechanism of B12b/DDTC-induced cytotoxicity in human MCF-7 cancer cell lines involves mitochondrial oxidative stress, activation of MPT pore and Ca2+ overload. Support of Funding Information: Russian Science Foundation № 19-75-20145 Citation Format: Marina E. Solovieva. Vitamin B12 augments Ca2+ dependent cytotoxicity of diethyldithiocarbamate through excessive ER-vacuolization, oxidative stress and mitochondrial depolarization in MCF-7 cells in culture [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2032.

Abstract 2356: Cyclin D1 mediated exosomes are enriched for pro-oncogenic miRNAs and promote cancer stem cell expansion

Abstract miRNAs are a class of endogenous noncoding small RNAs that participate in diverse biological processes through base-pairing to the complementary sequence in 3' untranslated region (3' UTR) of mRNA Specific miRNA have been associated with the initiation, progression and therapy resistance and regulate immunological processes including the activation and ageing of innate and adaptive immune cells. Immuno-miRs, including. miR-21 and miR-93, regulate immune function in the tumor microenvironment by binding to the 3′-UTR region of target mRNAs, or by binding to- and activating Toll-Like Receptor 8 (TLR8) in surrounding cells of the immune system. The cyclin D1 (CCND1) gene, which encodes the regulatory subunit of a holoenzyme that phosphorylates and inactivates the Rb protein, is overexpressed in up to 50% of human breast cancers. Cyclin D1, governs the expression, processing, the secretion and the relative proportion of secreted non-coding RNA subtypes (miRNA, rRNA, tRNA, CDBox, scRNA, HAcaBox. scaRNA, piRNA) in human breast cancer and the secretion of piRNA. Exosomes, which consist of a lipid bilayer membrane surrounding a small cytosol, contain constituents of their cell of origin, including mRNA and miRNA. Exosomes in the tumor microenvironment, participate in therapy resistance, cancer metastasis and the metastatic niche. Herein, knock down of cyclin D1 in MCF-7 cells with tet-inducible cyclin D1 shRNA decreased MCF-7 cell proliferation and mammosphere formation. Exosomes isolated from wild-type and cyclin D1 knock-down MCF-7 cell culture media, showed that exosomes from cyclin D1 knock down cells dramatically decreased proliferation of both wild-type and cyclin D1 knock-down cells, whereas the wild-type exosomes partially reversed the cyclin D1 knock down effects. Cyclin D1 knock-down exosomes decreased mammosphere formation of wild-type MCF-7 cells and wild-type exosomes increased mammosphere formation of cyclin D1 knock-down cells. miRNA profiles within the exosomes, showed that cyclin D1 regulated the abundance of miRNA within exosomes. The pattern of miRNA within exosomes regulated by cyclin D1 were enriched for miRNA governing mesenchymal epithelial transition and the induction of stem cell function. The most enriched miRNA in cyclin D1 wild-type exosomes was hsa-mir-21 which was recognized tumor promoting function, whereas the most enriched miRNAs in cyclin D1 knock-down exosomes were has-mir-16 and has-mir-92a, both of which were shown as tumor suppressor in breast cancer cells. The immune-miR miR-21 represented &amp;gt;85% of the cyclin D1-induced exosome miRNA transcripts, The cyclin D1-mediated miRNA exosome content may contribute to tumor initiation and progression through heterotypic expansion of cancer stem cells. Citation Format: Xuanmao Jiao, Chongwen Xu, Lifeng Tian, Zhao Zhang, Anthony W. Ashton, Zhiping Li, Richard G. Pestell. Cyclin D1 mediated exosomes are enriched for pro-oncogenic miRNAs and promote cancer stem cell expansion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2356.

Increased expression of autophagy and Nrf2-dependent signaling pathway genes by new monophenolic antioxidants depends on their structure

Under certain conditions, both activators and inhibitors of Nrf2-dependent signaling and autophagy can serve as potential agents for the prevention, treatment, and maintenance therapy of cancer, as well as overcoming chemoresistance. We are developing a panel of structurally related synthetic monophenolic compounds, in addition to a direct antioxidant effect, having the ability to affect intracellular signaling pathways and processes, including the above mentioned. Aim of the study was to investigate the relationship between its structure and the ability to influence the expression of autophagy and Nrf2 pathway genes.Material and methods. Five original hydrophilic phenolic structurally related compounds were synthesized; the prototypical inductor of the Keap1/Nrf2/ARE system, monosubstituted phenol tert-butylhydro-quinone (tBHQ), served as a reference substance. We used cultures of murine Lewis lung carcinoma (LLC) and human breast adenocarcinoma MCF-7 cells, with the test compounds at a concentration of 20 gM cells were incubated for 24 hours. Changes in mRNA expression of Nrf2, Nqo1, LC3b, and Sqstm1 genes encoding Nrf2, NAD(P)H:quinone oxidoreductase-1, LC3B, p62/sequestosome-1, respectively, were determined by TaqMan real-time PCR.Results and discussion. Structurally related synthetic monophenols effectively induced the Keap1/Nrf2/ARE system in LLC and MCF-7 cells, increasing the expression of the Nrf2-driven Nqo1 gene, and ambiguously affected the transcription of genes of proteins mediating autophagy: the content of mRNA of the LC3B protein did not change, while the expression of mRNA of the p62 protein, which affects the operation of both systems (processes), is increased only by the action of ortho-monosubstituted phenol with an «active» sulfur atom in the para-propyl substituent TS-13 and its structural analogue TS-12 with a para-substituent shortened by one methylene. It can be assumed that the observed effect either indicates the ability of TS-13 and TS-12 both to induce the Keap1/Nrf2/ARE system and to stimulate autophagy, or reflects a compound-mediated mutually reinforcing cross-activation of these processes.

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

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 anti-breast 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.

Building Block Symmetry Relegation Induces Mesopore and Abundant Open-Metal Sites in Metal–Organic Frameworks for Cancer Therapy

Building Block Symmetry Relegation Induces Mesopore and Abundant Open-Metal Sites in Metal–Organic Frameworks for Cancer Therapy

Epigenetic modulators combination with chemotherapy in breast cancer cells.

Despite the concerning adverse effects on tumour development, epigenetic drugs are very promising in cancer treatment. The aim of this study was to compare the differential effects of standard chemotherapy regimens (FEC: 5-fluorouracil plus epirubicine plus cyclophosphamide) in combination with epigenetic modulators (decitabine, valproic acid): (a) on gene methylation levels of selected tumour biomarkers (LINE-1, uPA, PAI-1, DAPK); (b) their expression status (uPA and PAI-1); (c) differentiation status (5meC and H3K27me3). Furthermore, cell survival as well as changes concerning the invasion capacity were monitored in cell culture models of breast cancer (MCF-7, MDA-MB-231). A significant overall decrease of cell survival was observed in the FEC-containing combination therapies for both cell lines. Methylation results showed a general tendency towards increased demethylation of the uPA and PAI-1 gene promoters for the MCF-7 cells, as well as the proapoptotic DAPK gene in the treatment regimens for both cell lines. The uPA and PAI-1 antigen levels were mainly increased in the supernatant of FEC-only treated MDA-MB-231 cells. DAC-only treatment induced an increase of secreted uPA protein in MCF-7 cell culture, while most of the VPA-containing regimens also induced uPA and PAI-1 expression in MCF-7 cell fractions. Epigenetically active substances can also induce a re-differentiation in tumour cells, as shown by 5meC, H3K27me3 applying ICC. SIGNIFICANCE OF THE STUDY: Epigenetic modulators especially in the highly undifferentiated and highly malignant MDA-MB-231 tumour cells significantly reduced tumour malignancy thus; further clinical studies applying specific combination therapies with epigenetic modulators may be warranted.

IC50 and Cell Viability of Combination of Ethanol Extract Moringa oleifera Leave (EEMo) and Ethanol Extract Carica papaya Leave (EECp) on Breast Cancer Cells

This study aims to find out IC50 and Cell Viability of the combination of ethanol extract leaves of Moringa oleifera (EEMo) and ethanol extract leaves of Carica papaya (EECp) on the growth of MCF-7 breast cancer cells culture. It was conducted in a true experimental laboratory using post-test only control group design method. The study showed that the effect of extract combination for MCF-7 Cell by using series concentration like 0, 20, 40, 80 and 160 µg/mL for 48, 72, and 96 hours with a cell density of 5x103 after giving WST assay there is a decrease in the number of cell viability. Inhibition concentration of MCF-7 cell culture was also indicated by the IC50 value which was included in the very strong category with details of each extract combination with 1:1 comparison the IC50 value is 12.02 µg/mL. &#x0D; &#x0D; Keywords: extract combination; IC50; cell viability, MCF-7

Comparative Study of Lipophilic Statin Activity in 2D and 3Din vitro Models of Human Breast Cancer Cell Lines MDA-MB-231 and MCF-7.

IntroductionStatins are a type of drugs that are used to lower cholesterol level in blood. Since the early 1990s, it has been known that statins could be beneficial in cancer therapy. However, data remain controversial, especially regarding estrogen receptors status. Despite many studies in breast cancer models in vitro, the correlations of effects of separate statins in various model systems remain unclear.AimOur aim was to evaluate the anticancer activity of lovastatin, mevastatin, pitavastatin and simvastatin on different subtypes of human breast cancer (MDA-MB-231 and MCF-7 cell lines) in spatially different 2D and 3D cultures in vitro.Materials and MethodsCell viability was tested using MTT assay. Effect of statins on cell colony formation was evaluated by calculating breast cancer cell colony area and number. The effect on cell migration was estimated by “wound healing” assay. The activity of compounds in 3D cultures was evaluated by measuring the spheroid size changes during incubation.ResultsAmong the tested statins, pitavastatin had the greatest effect on the viability of breast cancer MDA-MB-231 and MCF-7 cell lines. The mevastatin and pitavastatin mostly decreased the MDA-MB-231 and MCF-7 cell colony formation. All statins at 90% of their estimated effective concentration (EC50) and simvastatin at 10% of its EC50 concentration suppressed the MCF-7 cells migration in “wound healing” assay. Only higher concentrations of mevastatin and pitavastatin slowed down the MDA-MB-231 cell migration. Statins showed different activity on 3D cell cultures growth. Lovastatin and simvastatin delayed the growth of MDA-MB-231 cell spheroids, while mevastatin and pitavastatin reduced the growth of MCF-7 spheroids.ConclusionStatins possess different anticancer activity in human breast cancer MDA-MB-231 and MCF-7 cell cultures. Pitavastatin and simvastatin showed the highest activity in most tested assays, especially against MCF-7 cell line.

The Influence of the Faradarmani Consciousness Field on the Survival and Death of MCF-7 Breast Cancer Cells: An Optimization Perspective

Mohammad Ali Taheri, the inventor of Faradarmani method believes beyond matter and energy fields, there are consciousness fields that can be accessed through connection of the consciousness of the subject of study to the cosmic consciousness network. This causes an optimization in a subject’s structure and function. Apart from the fact that most studies in the field of cancer are aimed at the death of cancer cells, the purpose of this study was to investigate the MCF-7 cancer cell survival and death according to their optimal nature under the influence of the Faradarmani consciousness field. For the purpose of this investigation, cellular viability was evaluated by MTT assay for 6-24 h under the influence of Faradarmani consciousness field. Consequently, cell cycle analysis was performed for evaluation of apoptotic cell death. Finally, the expression of Bcl2 and Bax genes in MCF-7 cells was evaluated as a validation of the obtained cellular scale results. Our findings demonstrate that the Faradarmani consciousness field treatment significantly induced cell proliferation in the MCF-7 cell culture in specific test conditions mediated by increasing cellular viability by about 18% in comparison to the control in a time-dependent manner. Moreover, the S-phase in the cell cycle, as a sign of cell population in the sample treated with Faradarmani was increased by 56% (up to 24h) in comparison to the control. Moreover, the results of the Realtime PCR reaction show that in cells treated with Faradarmani consciousness field, the Bax / Bcl2 message ratio decreased after 24 h (more than 1-fold negative control (untreated)) suggesting a higher cell survival and resistance to death. Considering the concepts of the consciousness fields according to Taheri, proliferation and survival appear to become optimized in MCF-7 cancer cell lines under influence of Faradarmani consciousness field.

Hybrid mesoporous silica-based nanocarriers for responsive drug release in cancerous cell line

In this work, the synthesis of organic–inorganic hybrids based on pH-responsive, poly(acrylic acid) (PAA) and thermoresponsive, poly(N-isopropyl acrylamide) (PNIPAM), grafted from RAFT agent-primed mesoporous silica nanoparticles (MSNs), has been studied. The confirmation of polymer grafting was obtained through several techniques, for example, FTIR spectroscopy, NMR spectroscopy, etc. The presence of C–H stretching vibrations from FTIR confirmed the presence of organic network in the inorganic MSNs. Further support appeared as notable resonances in 13C solid-state NMR. The resonance at 175 ppm from C=O group for PAA-grafted MSNs, and at 170–172 ppm from C=O group for PNIPAM-grafted MSNs confirmed the grafting of polymer from the RAFT agent-primed MSNs. Morphological analysis for PAA- and PNIPAM-grafted MSNs was performed using FESEM and TEM. The images manifested spherical shape for isobutyric acid group, and short rod shape for phenyl ethyl group-containing RAFT agent-primed MSNs. Subsequently, for assessing their effectiveness as drug delivery vehicle, the anti-cancerous drug doxorubicin hydrochloride (Dox) was loaded into the MSNs. An effective loading in the range of 50–55% in case of PAA-grafted and 49–61% in case of PNIPAM-grafted MSNs (at pH 7.4, 25 °C) was observed. Subsequently release efficiencies for these Dox-encapsulated MSNs were studied at varying pH, temperature and time. The treatment of cultured MCF-7 cell lines by the control MSNs and polymer-grafted MSNs revealed that they are non-toxic. However, when the MSNs were Dox-loaded, the PAA-grafted ones demonstrated higher cytotoxicity than the PNIPAM-grafted MSNs at equivalent dose at pH 7.4 and 37°C. Together with other established features, we show that the polymer-grafted MSNs studied in this work can be utilized as an efficient drug delivery system for different therapeutic applications.