Human Cell Line MCF-7 Research Articles

Synthesis, Anticancer Activity, Docking Calculations and Hydrolytic Stability Studies of Bioconjugates of Monofluorenated Analogue of BIM- 23052.

The fight against cancer has started since its discovery and has not subsided to nowadays. Currently, the hybrid molecules have become a promising alternative to the standard chemotherapeutics for the treatment of multi-causal diseases, including cancers. Herein, we report the synthesis, biological evaluation, mathematical docking calculations and hydrolytic stability of the new bioconjugates of monofluorinated analogues of BIM-23052, containing second pharmacophore naphthalimide, caffeic acid or the tripeptide Arg-Gly-Asp. All new molecules are obtained using standard peptide synthesis on solid support. Anticancer potential is studied against a panel of tumor cell lines included human mammary carcinoma cell lines MCF-7 (ER+, PR+ and Her-2-); MDA-MB-231 (ER-, PR- and Her-2-), as well as cell lines BALB 3T3 (mouse embryonic fibroblasts) and MCF-10A (human breast epithelial cell line). The IC50 values found in the MCF-10A cell line assay were used to calculate the selective index (SI). The highest SI relative to MCF-7, with a value of 2.62 is shown by the compound Npht- Gly-D-Phe-Phe(4-F)-Phe-D-Trp-Lys-Thr-Phe-Thr-NH2. In MCF-10 cells, the weakest antiproliferative effect was caused by the same compound (IC50 = 622.9 ± 23.91 μM), which makes this analogue a good candidate for the new anticancer medical drug. Unfortunately, the hydrolytic stability studies reveal that this bioconjugate is the most unstable of hydrolysis under physiological conditions in the body. Even with lower anticancer activity and selectivity in comparison with Npht-Gly-DPhe- Phe(4-F)-Phe-D-Trp-Lys-Thr-Phe-Thr-NH2, the compound Arg-Gly-Asp-D-Phe-Phe(4-F)-Phe- D-Trp-Lys-Thr-Phe-Thr-NH2 is the best candidate between three investigated bioconjugates for practical application due to combination of activity and stability profiles. Mathematical docking calculation also reveals that synthesized bioconjugates show selectivity according to different somatostatin receptors on the surface of different cell lines.

In silico and in vitro studies on the anti-cancer activity of artemetin, vitexicarpin and penduletin compounds from Vitex negundo

Vitex negundo L. (V. negundo) is one of the important medicinal and anticancer enhancer herbs. This plant is commonly used in the preparation of traditional drugs to treat numerous diseases. Inspired by the medicinal properties of this plant, the current study aimed to investigate antiproliferative potential and the primary molecular mechanisms of the apoptotic induction against human HepG2 and MCF-7 cell lines, by pure compounds isolated from targeted fractions of V. negundo which were characterized by NMR, FTIR and HRMS analysis and identified as artemetin (FLV1), vitexicarpin (FLV2), and penduletin (FLV3) compounds. The FLV1, FLV2, and FLV3 compounds were evaluated for the antiproliferative potential against HepG2 and MCF-7 cell lines by cell viability assay and exhibited IC50 values of 2.3, 23.9 and 5.6 µM and 3.9, 25.8, and 6.4 µM, respectively. In addition, those compounds increased the level of reactive oxygen species production, induced cell death occurred via apoptosis, demonstrated by Annexin V-staining cells, contributed significantly to DNA damage, and led to the activation of caspase3/caspase8 pathways.Additionally, molecular docking was also conducted to rationalize the cancer cells inhibitory and to evaluate the ability of the FLV1, FLV2, and FLV3 compounds to be developed as good drug candidates for cancers treatment.

In vitro anticancer activity of Semecarpus anacadium on MCF-7 human breast cancer cells

Semecarpus anacardium (SA) is a plant native to India that is eaten as a fresh vegetable on a daily basis. The effects of SA therapy on human breast cancer MCF-7 cell lines were investigated in this study. Maceration with various solvents was used to prepare the crude extract of flavonoid fraction from SA bark, and fractionation was done using TLC and column chromatography techniques. MTT assay was used to assess the effects of the isolated flavonoid fraction of SA bark on cell viability. SA decreased MCF-7 cancer cell viability in a concentration-dependent manner, with IC50 values of 24.18+1.20 μg/mL and 12.08+1.18 μg/mL after 24 and 48 hours of treatment, respectively. Finally, SA flavonoid fraction effectively reduced MCF-7 cells, implying that SA flavonoid fraction has anticancer potential against MCF-7 cells and may be useful in the prevention and treatment of breast cancer.

Syntheses and high selective cytotoxicity of dehydroabietylamine C-ring nitration derivatives

To find more effective anticancer agents, a series of novel dehydroabietylamine (DA) derivatives were synthesized, focusing on C-ring nitro modifications and C-18 imide introduction. Their cytotoxic activities against human tumor cell line HeLa (cervix), MCF-7 (breast), A549 (lung), HepG2 (liver), and nonmalignant cell line HUVEC (umbilical vein) in vitro were screened. The C-18 imide heterocyclic compounds 1, 2, and C-ring 14-nitro substituted 14 exhibited moderate to good cytotoxic activities and significant selectivity towards malignant cell lines. More importantly, they were significantly less cytotoxic to nonmalignant cells (HUVEC) than the parent compound and positive control doxorubicin hydrochloride (DOX). Meantime the mechanism of cytotoxicity of DA derivatives was studied. Annexin V-FITC/PI double-staining analysis suggested that cytotoxicity of compounds 2 and 14 was associated with early apoptosis induction. The interaction between compounds and DNA (herring sperm DNA) was studied using absorption spectral analysis and ethidium bromide (EB) fluorescence displacement experiments, the results exhibited that the binding of the compound to DNA was in the intercalative mode. The structure-activity relationship discussion implied that introduction of the nitro-group, especially the 14-nitro group, can significantly improve the cytotoxicity of dehydroabietylimide compounds. The relatively high cytotoxicity and significant high selectivity of compounds 2 and 14 indicated that they were particularly noteworthy. NO released amounts indicated that the amounts of NO released by the compounds bearing nitro-group were quite well associated positive correlation with their cytotoxic activity, which provide a new strategy for structure design of DA anticancer agents in the future.

Citric acid-mediated green synthesis of selenium nanoparticles: antioxidant, antimicrobial, and anticoagulant potential applications.

Using microwave technique in the presence of citric acid, selenium nanoparticles (SeNPs) were fabricated. The morphological characteristics revealed that the spherical SeNPs with diameters ranging from 10.5 to 20 nm aggregated spherical shapes with sizes ranging from 0.67 to 0.83 mm. Moreover, the antioxidant efficacy was assessed by the DPPH radical scavenging test, which depicted that green-prepared nanoparticle at a 106.3 mg/mL dosage had the maximum scavenging capacity (301.1 ± 11.42 mg/g). Otherwise, with nanoparticle concentrations of 500 mg/ml, in vitro cell viability of SeNPs through human breast cancer MCF-7 cell lines was reduced to 61.2 ± 2.2% after 1 day of exposure. The antibacterial activity was tested against G-negative Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli), G-positive bacteria Bacillus subtilis (B. subtilis), and Staphylococcus aureus (S. aureus), which demonstrated that SeNPs had little activity against S. aureus. Still, it had the highest activity against E. coli, with a zone of inhibition (ZOI) of 25.2 ± 1.5 mm compared to 16.0 ± 0.6 mm for the standard antibiotic. Most notably, biogenic SeNPs have anticoagulant activities using activated partial thromboplastin time (aPTT) assessment. Based on previous findings, SeNPs can be used in medical aid and their cell viability, antioxidant, anticoagulant, and effects on bacteria.

3D Bioprinted Artificial Breast Cancer Tumor Disks: A New Tool for Understanding Intra‐tumor Heterogeneity?

Recently the UWRF Tissue and Cellular Innovation Center and the Century College Biofabrication Program have engaged in a series of new collaborative research projects using bioprinting technology and methods to model and study in‐vitro 3D artificial tumor and stromal tissues. In this presentation, we report on results obtained from efforts to bioprint cancer disks using the human breast adenocarcinoma cell line MCF‐7. This breast cancer cell line is considered to be one of the “gold standard” cultures to study this type tumor. In extensive previous studies in our lab using other 3D modeling methods, we have established a robust library of experience and data with these cells and artificial tissue constructs. In these new studies, bioprinting methods were applied to establish tumor structures which initially contained specific cell numbers per unit volume as well as relatively mono‐cellular distribution patterns. Both of these characteristics had been very difficult to predictably generate and control in earlier studies using precast or molded hydrogels made of chitosan and hyaluronic acid or decellularized natural marine sponge ECM. In this project, two distinct bioinks were tested including one called GelMa, a gelatin/methacrylate hydrogel and another called GelMa‐C which is basically the same, but with nanocellulose added to enhance the structural stability of constructs during printing. While both bioinks generated structures and tissues, the GelMa bioink specifically generated relatively transparent tumor structures that provided a unique window into the cell interactions and eventual early development of these artificial tumor tissues. Using a Cellink “Inkcredible” model printer loaned to the TCIC by Century College for the duration of this project, we printed tumor disks with a diameter of 5 mm and thickness of 2.5 mm and then cultured these in 6 well plates for up to 3 months. These breast cancer disks displayed features and tissue progression similar to that seen with the other methods used in our earlier work, while also providing new and unique insights into the interior artificial tumor tissue development in real‐time. These new observations also demonstrated a unique cell/tissue behavior in the resulting constructs that we hypothesize may directly address the origins of intra‐tumor heterogeneity of various cell subpopulations and their relative contributions to the overall tumor development. Future studies are planned to further evaluate and qualify these observations and to apply this new 3D artificial tissue tool to further elucidate cancer biology as a whole.

Effects of tumor-derived extracellular vesicles on T cell fate and function

Abstract Immunosuppressive microenvironments created by malignant tumors represent a major obstacle for effective anti-tumor immunity. There are multiple immunosuppressive mechanisms operative in the tumor microenvironment (TME) that interfere with T cell function. Extracellular vesicle (EV) in tumor microenvironment is an important way employed by tumor cells to control T cell fate and function. However, the mechanisms responsible for EV-mediated T cell suppression and dysfunction are still largely unknown. Here, we purified EVs from human melanoma A375 and breast cancer MCF7 cell lines and then explored EV impacts on T cell function and fate in anti-tumor immunity. We found that tumor derived EVs can suppress T cell proliferation and induce T cell senescence, which is a novel and fundamentally important phenomenon that contributes to tumor-induced immunosuppression in the TME. Mechanistically, tumor derived EVs promoted DNA damage responses and cell cycle arrest of T cell that induces T cell senescence. Blockage of DNA damage signaling in T cells via the specific inhibitor KU55933 can prevent T cell senescence and recover T cell impaired effector functions. Importantly, we further performed in vivo studies to investigate that tumor-derived EVs can induce T cell suppression and senescence, which can also be reversed by specific signaling inhibitors or/and targeting signaling molecules via the CRISPR knockdown strategy. These studies facilitate a better understanding of the novel suppressive mechanism utilized by the TME and provide insights relevant to the development of effective strategies to reverse immune suppression for enhanced anti-tumor immunity. Supported by the grants from the Melanoma Research Alliance and National Institutes of Health.

Targeting Cell Death Mechanism Specifically in Triple Negative Breast Cancer Cell Lines.

Triple negative breast cancer (TNBC) is currently associated with a lack of treatment options. Arsenic derivatives have shown antitumoral activity both in vitro and in vivo; however, their mode of action is not completely understood. In this work we evaluate the response to arsenate of the double positive MCF-7 breast cancer cell line as well as of two different TNBC cell lines, Hs578T and MDA-MB-231. Multimodal experiments were conducted to this end, using functional assays and microarrays. Arsenate was found to induce cytoskeletal alteration, autophagy and apoptosis in TNBC cells, and moderate effects in MCF-7 cells. Gene expression analysis showed that the TNBC cell lines’ response to arsenate was more prominent in the G2M checkpoint, autophagy and apoptosis compared to the Human Mammary Epithelial Cells (HMEC) and MCF-7 cell lines. We confirmed the downregulation of anti-apoptotic genes (MCL1, BCL2, TGFβ1 and CCND1) by qRT-PCR, and on the protein level, for TGFβ2, by ELISA. Insight into the mode of action of arsenate in TNBC cell lines it is provided, and we concluded that TNBC and non-TNBC cell lines reacted differently to arsenate treatment in this particular experimental setup. We suggest the future research of arsenate as a treatment strategy against TNBC.

Optimization of pre-analytical and analytical steps for DNA and RNA analysis of fresh cytology samples.

Different cytology preparations can be used for molecular diagnostics, however the influence of pre-analytical and analytical steps on the results are not yet well defined. We aimed to determine optimal steps for efficient extraction of DNA and RNA from fresh cells for molecular diagnostics. MCF7 and FaDu human cell lines, were used as a model to determine fresh cells storage conditions (temperature: 25°C, 4°C, -20°C, -80°C; duration: 0h, 4h, 12 h, 24 h, 48 h) and optimal nucleic acids extraction method. Besides, the minimal number of total cells and minimal percentage of mutated cells needed for successful extraction of nucleic acids and subsequent determination of present mutation were evaluated. Extraction of nucleic acids using spin columns yielded the highest quantity and quality of nucleic acids. Isolation of nucleic acids was feasible in all storage conditions, however higher temperature and longer duration of fresh cells storage were associated with lower quality of isolated nucleic acids and similar quantification cycle of housekeeping genes. Successful molecular testing was feasible with least 104 cells, while specific mutation was detected in as low as 5% of mutated cells. Our cell line model, mimicking fresh cytology samples, showed that quantity of extracted either DNA or RNA declined with higher temperatures and longer duration of storage but regardless of the storage conditions, we successfully detected both housekeeping genes and mutated gene using qPCR.

Cytotoxic and antiproliferative potential of methanolic extract of Mallotus phillippensis in MCF-7 cell line

Mallotus philippensis is a shrub, used for anti-filarial, anti-bacterial, anti-inflammatory, anthelmintic and immune-regulatory purposes for centuries. In the current study, methanolic extract of M. phillippensis leaves was examined in vitro for cytotoxicity and apoptotic potential in MCF-7 breast cancer cell lines. Various biochemical tests used to investigate the phytochemical contents of the plant extract. Qualitative phytochemical analysis of M. phillippensis leaves revealed the presence of phenolic compounds, steroids, flavonoids, alkaloids, diterpenes and tannins. The MTT dye reduction test was used to assess the cytotoxicity of the methanol extract of M. phillippensis. MCF-7 cells were grown in RPMI medium supplemented with 10% serum and 1% antibiotic and antimycotic solution. M. phillippensis leaf extract reduced cell viability in a dose-dependent manner, with a half maximal inhibitory concentration (IC50) of 190 g/mL. Acridine orange/ Ethidium bromide and fluoroprobe, benzimidazol-carbocyanine iodide 5, 5’, 6, 6’-tetrachloro-1, 1’, 3,3’-tetra ethyl (JC-1) staining. were used to examine the influence of the extract on the development of apoptotic characteristics in MCF-7 cells. The AO/EB staining revealed apoptotic cells with orange-red fluorescence in extract-treated cells, which increased in a dose-dependent manner whereas control cells showed green fluorescence. Control cells stained with JC-1 dye fluoresced redorange, whereas cells treated with extract showed red-green fluorescence whose intensity changed in a dose-dependent manner. The methanolic extract of M. phillippensis leaves showed invitro cytotoxic potential as well as induced apoptotic cell death in human breast cancer MCF-7 cell lines and may be studied further to generate a potent anti- cancer drug.

Anti-cancer Drug Anlotinib Promotes Autophagy and Apoptosis in Breast Cancer.

Anlotinib, a multi-target tyrosine kinase inhibitor, has significant anti-cancer effects on breast cancer (BC), lung cancer, colon cancer and ovarian cancer, but its mechanism has not been investigated in BC. The cell viability and growth of human non-triple negative BC cell line MCF-7 and triple negative BC cell line MDA-MB-231 with the treatment of anlotinib were tested by Cell Counting Kit-8 (CCK-8) assay and Ki67 staining. The alteration of genes related to apoptosis and autophagy were investigated by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR), western blots and immunocytochemistry (ICC). Cell apoptosis was valued by TUNEL staining and flow cytometry. Further, mouse breast tumour cell lines AT-3 cells were subcutaneously injected into C57BL/6 mice, and the effect of anlotinib intragastrically on tumour growth in vivo was examined. We found that anlotinib suppressed the cell viability and depressed Ki67 staining in MCF-7 and MDA-MB-231 cell lines. Besides, the drug also enhanced cell autophagy and apoptosis of MCF-7 and MDA-MB-231 cell lines, which could be rescued by autophagy inhibitors wortmannin (wort) and 3-methyladenine (3-MA), and BECN1 knockdown. Furthermore, Akt/GSK-3α pathway was inactivated by anlotinib treatment, while rescued by wort, 3-MA and silencing of BECN1 in the MCF-7 or MDA-MB-231 cells. We also found that anlotinib inhibited implanted tumour growth of BC in syngeneic mice. Our study demonstrated that anlotinib inhibited breast cancer cell growth in vitro and in vivo. Anlotinib promoted cell apoptosis and inactivated Akt/GSK-3α pathway of BC cells by inducing cell autophagy. It indicated that anlotinib may be an effective new drug for BC treatment.

ADAPTOR PROTEIN Ruk/CIN85 PARTICIPATES IN THE METABOLIC CONTROL OF HUMAN BREAST ADENOCARCINOMA MCF-7 CELLS

Aim. To determine the role of Ruk/CIN85 in the control of breast adenocarcinoma cells metabolism, we performed systemic analysis of the activity levels/content of key enzymes/components of glycolysis and oxidative phosphorylation using as a model the weakly invasive human breast adenocarcinoma MCF-7 cell line (Mock); and its sublines with stable overexpression (G4 subline) and reverse down-regulation (G4vir subline) of the adaptor protein. Materials and methods. MCF-7 cells were cultured in the complete DMEM medium under standard conditions. Enzymes activity, content of metabolites and protein in cell extracts and the conditioned cell culture medium were estimated by spectrophotometric and fluorometric assays. Results. First of all, biochemical indexes of aerobic glycolysis, activity levels of some key glycolytic enzymes and metabolites were evaluated. A significant increase in the activity of these enzymes, aldolase A (ALDOA) and lactate dehydrogenase A (LDHA), was found in G4 cells compared to Mock by 1.3 and 1.6 times, respectively. In addition, in the conditioned medium of G4 cells, an increase in lactate content by 1.5 times compared with the control was found, which corresponded to a change in LDHA activity. Knockdown of Ruk/CIN85 expression level in G4 subline resulted in a significant decrease of these parameters compared to G4 cells, ALDOA – 4 times, LDHA - 1.4 times, and lactate production - 2.5 times. It should be noted that in G4vir cells, LDHA activity returned to level of control cells, while ALDOA activity and lactate content additionally decreased by 3 times and 1.6 times, respectively. Therefore, the observed changes in the intensity of glycolysis in MCF-7 sublines positively correlate with the expression level of adaptor protein studied. To assess the metabolic status of mitochondria, the level of activity of the Krebs cycle enzyme, NAD-dependent malate dehydrogenase (MDH2), the catalyst of last stage of the cycle, was determined. A 2-fold decrease in MDH2 activity was found in the MCF-7 G4 subline relative to control Mock cells, as well as an increase in this index by 2.4 times in G4vir cells to control values. Unlike glycolysis, we observed the opposite pattern with respect to the intensity of Krebs cycle reactions depending on the expression level of Ruk/CIN85. Conclusions. Use of limited proteolysis technique as the source of additional information for computer modeling allowed us to propose an improved model of 3D-structure of fibrinogen αC-regions. This model takes into account the behavior of αC-regions in the physiological condition and contributes to the general knowledge about fibrinogen structure.

Attachment of Chiral Functional Groups to Modify the Activity of New GPx Mimetics.

A series of new chiral benzisoselenazol-3(2H)-ones and their corresponding diselenides bearing an o-amido function substituted on the nitrogen atom with various aliphatic and aromatic moieties were synthesized. All derivatives representing pairs of enantiomers or diastereoisomers were obtained to thoroughly evaluate the three-dimensional structure–activity correlation. First, bensisoselenazol-3(2H)-ones were synthesized by reacting 2-(chloroseleno)benzoyl chloride with an appropriate enantiomerically pure amine. Then, the Se–N bond was cleaved by a reduction–oxidation procedure using sodium borohydride and then air oxidation to obtain the corresponding diselenides. All derivatives were tested as antioxidants and anticancer agents. In general, the diselenides were more reactive peroxide scavengers, with the highest activity observed for 2,2′-diselenobis[N-(1S,2S)-(-)-trans-2-hydroksy-1-indanylbezamide]. The most cytotoxic derivative towards human promyelocytic leukemia HL-60 and breast cancer MCF-7 cell lines was N-[(1S,2R)-(-)-cis-2-hydroksy-1-indanyl]-1,2-benzizoselenazol-3(2H)-one. The structure–activity relationship of the obtained organoselenium derivatives was discussed.

Design, synthesis of anticancer and anti-inflammatory 4-(1-methyl-1H-indol-3-yl)-6-(methylthio) pyrimidine-5-carbonitriles

A novel series of 4-(1-methyl-1H-indol-3-yl)-6-(methylthio) pyrimidine-5-carbonitriles (4a–i) was synthesized and evaluated for anticancer potential against cell lines for breast cancer. Compounds 4b, 4e, and 4h exhibited prominent cytotoxicity against human breast carcinoma MCF-7 cell line with GI50 of 2.0, 0.5, and 0.5 µM, respectively. Molecular docking study against EGFR tyrosine kinase could provide valuable insights into the plausible mechanism of action. The compounds could bind with significantly high binding affinity and their binding affinity scores could correlate well with the observed anticancer activity. Furthermore, compounds 4a, 4c, 4e, 4g, and 4i exhibited significant inflammatory activities as well which could expand the therapeutic domain of this novel series.

Arene-ruthenium(II) complexes with tetracyclic oxime derivatives: synthesis, structure and antiproliferative activity against human breast cancer cells

Six new arene-ruthenium(II) complexes containing two different η6-arene ligands – benzene and hexamethylbenzene, with indenoquinoxalinone oxime analogues (11H-indeno[1,2-b]quinoxalin-11-one oxime, 6H-indeno[1,2-b]pyrido[3,2-e]pyrazin-6-one oxime and 6–(hydroxyimino)indolo[2,1-b]quinazolin-12(6H)-one oxime (tryptanthrin-6-oxime)) as N,N’- chelating ligands are reported. The complexes were characterized by elemental analysis, IR, UV–VIS, 1H and 13C NMR spectroscopy and by single-crystal X-ray structure analysis. Complexes adopt a half-sandwich «piano-stool» geometry with oxime ligands in anionic form, the ruthenium(II) center coordinates one arene, one N,N-bidentate oximate and one chloride ligand. The computational analysis of non-covalent intermolecular interactions revealed weak attraction between the oxime oxygen atoms and the nearest hydrogen atoms of the oxime and the arene ligands. The cytotoxic activity of the complexes was evaluated against human breast cancer cell line MCF-7 and cisplatin-resistant human breast cancer cell line MCF-7CR as well as non-cancerous human breast epithelial cell line MCF10A. The cytotoxicity tests show micromolar IC50 values and tryptanthrin-6-oxime hexamethylbenzene-ruthenium(II) complex was found to exhibit the best antiproliferative activity among the studied compounds against the MCF-7 and MCF-7CR cell lines (IC50 9.0 ± 4.4 and 8.9 ± 1.5 µM, respectively).

Epigenetic study of early breast cancer (EBC) based on DNA methylation and gene integration analysis

Breast cancer (BC) is one of the leading causes of cancer-related deaths in women. The purpose of this study is to identify key molecular markers related to the diagnosis and prognosis of early breast cancer (EBC). The data of mRNA, lncRNA and DNA methylation were downloaded from The Cancer Genome Atlas (TCGA) dataset for identification of differentially expressed mRNAs (DEmRNAs), differentially expressed lncRNAs (DElncRNAs) and DNA methylation analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyzes were used to identify the biological functions of DEmRNAs. The correlation analysis between DNA methylation and DEmRNAs was carried out. Then, diagnostic analysis and prognostic analysis of identified DEmRNAs and DElncRNAs were also performed in the TCGA database. Subsequently, methylation state verification for identified DEmRNAs was performed in the GSE32393 dataset. In addition, real-time polymerase chain reaction (RT-PCR) in vitro verification of genes was performed. Finally, AC093110.1 was overexpressed in human BC cell line MCF-7 to verify cell proliferation and migration. In this study, a total of 1633 DEmRNAs, 750 DElncRNAs and 8042 differentially methylated sites were obtained, respectively. In the Venn analysis, 11 keys DEmRNAs (ALDH1L1, SPTBN1, MRGPRF, CAV2, HSPB6, PITX1, WDR86, PENK, CACNA1H, ALDH1A2 and MME) were we found. ALDH1A2, ALDH1L1, HSPB6, MME, MRGPRF, PENK, PITX1, SPTBN1, WDR86 and CAV2 may be considered as potential diagnostic gene biomarkers in EBC. Strikingly, CAV2, MME, AC093110.1 and AC120498.6 were significantly actively correlated with survival. Methylation state of identified DEmRNAs in GSE32393 dataset was consistent with the result in TCGA. AC093110.1 can affect the proliferation and migration of MCF-7. ALDH1A2, ALDH1L1, HSPB6, MME, MRGPRF, PENK, PITX1, SPTBN1, WDR86 and CAV2 may be potential diagnostic gene biomarkers of EBC. Strikingly, CAV2, MME, AC093110.1 and AC120498.6 were significantly actively correlated with survival. The identification of these genes can help in the early diagnosis and treatment of EBC. In addition, AC093110.1 can regulate SPTBN1 expression and play an important role in cell proliferation and migration, which provides clues to clarify the regulatory mechanism of EBC.

Gadolinium(III) Coordinated Theranostic Polymer for Proficient Sequential Targeting–Combinational Chemotherapy and T1 Weighted Magnetic Resonance Imaging

The combination therapy where a single delivery agent codelivers multiple drugs at the targeted site has become a promising strategy to achieve synergistic chemotherapeutic effect leading to the suppression of cancer cell drug resistance capability. Herein, we developed a theranostic polymer CP PG MT Gd that can sequentially target folate receptors (FR) onto cancer cell apical membrane at an early phase and exhibited late phase combinational chemotherapy through topoisomerase-I (TOPO-I) and dihydrofolate reductase (DHFR) inhibition synergistically, which resulted in inhibition of nucleic acid biosynthesis. This dual-acting self-targeted CP PG MT Gd polymer could effectively track the loaded chemotherapeutic agents (camptothecin–methotrexate) through MRI-tomography and eventually played a decisive role for subsequent dose-determination in real-time.A thorough comparative investigation on FR-targeting competence of CP PG MT Gd was executed with a folic acid (FOL) conjugated CP PG FL Gd (targeted system) and a nontargeted CP PG Gd polymers using human cervical cancer, HeLa (high FR-expression), and breast cancer MCF-7 (low FR-expression) cell lines. The incorporated Gd(III) chelate-complex in the CP PG MT Gd theranostic polymer displayed remarkable T1 weighted MRI-proficiency with longitudinal specific relaxivity constant, r1 = 21.85 mM–1 S–1. Therefore, the MRI-tracking proficiency and sequential targeting–chemotherapy switchable CP PG MT Gd theranostic system opened the way for precise multidrug codelivery at cancer site that might have attractive potential for diagnosis and decisive dose-determination in clinical implication.

Synthesis and Evaluation of 1,3,5-Triaryl-2-Pyrazoline Derivatives as Potent Dual Inhibitors of Urease and α-Glucosidase Together with Their Cytotoxic, Molecular Modeling and Drug-Likeness Studies.

In the present work, a concise library of 1,3,5-triaryl-2-pyrazolines (2a–2q) was designed and synthesized by employing a multistep strategy, and the newly synthesized compounds were screened for their urease and α-glucosidase inhibitory activities. The compounds (2a–2q) were characterized using a combination of several spectroscopic techniques including FT-IR, 1H NMR, 13C NMR, and EI-MS. All the synthesized compounds, except compound 2i, were potent against urease as compared to the standard inhibitor thiourea (IC50 = 21.37 ± 0.26 μM). These analogs disclosed varying degrees of urease inhibitory activities ranging from 9.13 ± 0.25 to 18.42 ± 0.42 μM. Compounds 2b, 2g, 2m, and 2q having IC50 values of 9.36 ± 0.27, 9.13 ± 0.25, 9.18 ± 0.35, and 9.35 ± 0.35 μM, respectively, showed excellent inhibitory activity as compared to standard thiourea (IC50 = 21.37 ± 0.26 μM). A kinetic study of compound 2g revealed that compound 2g inhibited urease in a competitive mode. Among the synthesized pyrazolines, the compounds 2c, 2k, 2m, and 2o exhibited excellent α-glucosidase inhibitory activity with the lowest IC50 values of 212.52 ± 1.31, 237.26 ± 1.28, 138.35 ± 1.32, and 114.57 ± 1.35 μM, respectively, as compared to the standard acarbose (IC50 = 375.82 ± 1.76 μM). The compounds (2a–2q) showed α-glucosidase IC50 values in the range of 114.57 ± 1.35 to 462.94 ± 1.23 μM. Structure–activity relationship revealed that the size and electron-donating or -withdrawing effects of substituents influenced the activities, which led to the urease and α-glucosidase inhibiting properties. Compound 2m was a dual potent inhibitor against urease and α-glucosidase due to the presence of 2-CF3 electron-withdrawing functionality on the phenyl ring. To the best of our knowledge, these synthetic compounds were found to be the most potent dual inhibitors of urease and α-glucosidase with minimum IC50 values. The cytotoxicity of the compounds (2a–2q) was also investigated against human cell lines MCF-7 and HeLa. Compound 2l showed moderate cytotoxic activity against MCF-7 and HeLa cell lines. Moreover, in silico studies on most active compounds were also performed to understand the binding interaction of most active compounds with active sites of urease and α-glucosidase enzymes. Some compounds exhibited drug-like characteristics due to their lower cytotoxic and good ADME profiles.

In Vitro Investigation of the Antioxidant and Cytotoxic Potential of Tabernaemontana ventricosa Hochst. ex A. DC. Leaf, Stem, and Latex Extracts

Tabernaemontana ventricosa (Apocynaceae) a latex-bearing plant is used in traditional medicine for its therapeutic benefits in reducing fever and hypertension and wound healing. Due to limited information on the plant’s pharmacological activities, this study aimed to investigate the antioxidant potential of the leaf, stem, and latex extracts of T. ventricosa, using the Folin-Ciocalteu (total phenolics), aluminum chloride colorimetric (total flavonoids), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The cytotoxic activity was evaluated in the human HEK293 (embryonic kidney), HeLa (cervical carcinoma), and MCF-7 (breast adenocarcinoma) cell lines using the MTT assay. The latex extracts possessed the highest total phenolic content (115.36 ± 2.89 mg GAE/g), followed by the stem hexane extracts (21.33 ± 0.42 mg GAE/g), the chloroform leaf (7.89 ± 0.87 mg GAE/g), and the chloroform stem (4.69 ± 0.21 mg GAE/g) extracts. The flavonoid content was substantially high ranging from 946.92 ± 6.29 mg QE/g in the stem hexane, 768.96 ± 5.43 mg QE/g in the latex, 693.24 ± 4.12 mg QE/g in the stem chloroform, and 662.20 ± 1.00 mg QE/g in the leaf hexane extracts. The DPPH assays showed the highest percentage of inhibition at 240 µg/mL, for the stem hexane (70.10%), stem methanol (65.24%), and stem chloroform (60.26%) extracts, with their respective IC50 values of 19.26 µg/mL (stem hexane), 6.19 µg/mL (stem methanol), and 22.56 µg/mL (stem chloroform). The FRAP assays displayed minimal inhibition ranging from 4.73% to 14.40%, except for the latex extracts which displayed moderate inhibition at 15 µg/mL (21.82%) and substantial inhibition at 240 µg/mL (98.48%). The HeLa and MCF-7 cell lines were the most sensitive to the extracts, with the hexane, chloroform, and methanol leaf and stem, and latex extracts significantly affecting the percentage cell survival. Overall, the various parts of T. ventricosa exhibited strong antioxidant activity correlating to its cytotoxicity. Further studies should focus on the isolation of specific antioxidant compounds that could be investigated for their anticancer potential.

Antimicrobial and anticancer activities of Sesbania grandiflora mediated Cadmium oxide nanoparticles

ABSTRACT Cadmium oxide (CdO) nanoparticles have been biosynthesized via Sesbania grandiflora leaf extract and characterized using different analytical techniques. XRD analysis indicated FCC structure with average crystallite size being 45.77 nm. Morphological analyses involving SEM and HR-TEM confirmed the cubic crystalline nature of the prepared samples. The presence of Cd-O bond was ascertained using FTIR technique while the presence of the elements Cd and O was ensured using SEM-EDS. The synthesized CdO nanoparticles have exhibited good Antibacterial activity against both gram positive and gram negative bacteria. In the case of Antifungal studies, Aspergillus niger showed maimum sensitivity as compared to Aspergillus flavus. The bioinspired CdO nanoparticles have shown better Anticancer activity against MCF-7 and HeLa Human cell lines. Therefore, it is concluded that the Sesbania grandiflora leaf extract mediated CdO nanoparticles may serve as a potential candidate for a wide range of biomedical applications.