scholarly journals Multidimensional Profiling of Drug-Treated Cells by Imaging Mass Cytometry

2019 ◽  
Author(s):  
Alexandre Bouzekri ◽  
Amanda Esch ◽  
Olga Ornatsky
Keyword(s):  
Morphological Changes ◽  
Single Cells ◽  
Quantitative Information ◽  
Mass Cytometry ◽  
Microtubule Depolymerization ◽  
Ki 67 ◽  
Pairwise Correlation ◽  
Cellular Markers ◽  
Mcf 7

AbstractIn pharmaceutical research, high-content screening is an integral part of lead candidate development. Drug responsein vitroover 40 parameters including biomarkers, signaling molecules, cell morphological changes, proliferation indexes and toxicity in a single sample could significantly enhance discovery of new therapeutics. As a proof of concept, we present a workflow for multidimensional Imaging Mass Cytometry™ (IMC™) and data processing with open source computational tools. CellProfiler was used to identify single cells through establishing cellular boundaries, followed by histoCAT™ (histology topography cytometry analysis toolbox) for extracting single-cell quantitative information visualized as t-SNE plots and heatmaps. Human breast cancer-derived cell lines SKBR3, HCC1143 and MCF-7 were screened for expression of cellular markers to generate digital images with a resolution comparable to conventional fluorescence microscopy. Predicted pharmacodynamic effects were measured in MCF-7 cells dosed with three target-specific compounds: growth stimulatory EGF, microtubule depolymerization agent nocodazole and genotoxic chemotherapeutic drug etoposide. We show strong pairwise correlation between nuclear markers pHistone3S28, Ki-67 and p4E-BP1T37/T46in classified mitotic cells and anti-correlation with cell surface markers. Our study demonstrates that IMC data expands the number of measured parameters in single cells and brings higher-dimension analysis to the field of cell-based screening in early lead compound discovery.

scholarly journals In vitro/ In vivo Electrochemical Detection of Pt(II) Species

2021 ◽  
Author(s):  
Alexander Vaneev ◽  
Petr Gorelkin ◽  
Olga Krasnovskaya ◽  
Roman Akasov ◽  
Daniil Spector ◽  
...  
Keyword(s):  
Electrochemical Detection ◽  
Single Cells ◽  
Electrochemical Technique ◽  
Breast Adenocarcinoma ◽  
Tumor Spheroids ◽  
Tumor Bearing ◽  
Advanced Tumor ◽  
Mcf 7

The biodistribution of chemotherapy compounds within tumor tissue is one of the main challenges in the development of antineoplastic drugs, and novel techniques for simple, non-expensive, sensitive, and selective detection of various analytes in tumors are of great importance. In this paper we propose the use of platinized carbon nanoelectrodes (PtNE) for electrochemical detection of platinum-based drugs in various biological models, including single cells and tumor spheroids in vitro, and inside solid tumors in vivo. We have demonstrated quantitative direct detection of Pt(II) in breast adenocarcinoma MCF-7 cells treated with cisplatin and cisplatin-based DNP prodrug. To realize the potential of this technique in advanced tumor models, we measured Pt(II) in 3D tumor spheroids in vitro and tumor-bearing mice in vivo. The concentration gradient of Pt (II) species correlated with the distance from the sample surface in MCF-7 tumor spheroids. We then performed detection of Pt(II) species in tumor-bearing mice treated intravenously with cisplatin and DNP. We found that there was deeper penetration of DNP in comparison to cisplatin. This research demonstrates a novel minimally invasive, real-time electrochemical technique for the study of platinum-based drugs.

scholarly journals Synthesis of Chromene 4 Aldehyde and 6-Phenyl-6h-Chromeno [4,3-B] Quinoline Derivatives as Anticancer and Apoptosis Inducing Agents

2021 ◽  
Vol 9 (10) ◽  
pp. 1764-1772
Author(s):  
Rizuana Sultana
Keyword(s):  
Cell Lines ◽  
Morphological Changes ◽  
Quinoline Derivatives ◽  
Significant Extent ◽  
Diverse Range ◽  
Synthetic Strategy ◽  
Induced Apoptosis ◽  
Apoptosis Inducing Agents ◽  
Mcf 7

Abstract: A series of novel chromene 3-aldehyde and quinoline derivatives have been synthesized using diversely substituted nitroarenes in the presence of In, dil. HCl, H2O (reductive amination) and evaluated for in vitro cytotoxic activity in three different cancer cell lines (MDA-MB-453, MCF-7, A549 and PC3). The synthetic strategy utilized to access these hybrids is operationally simple and works with great substrate scope. Interestingly, compound 6i was induced apoptosis to a significant extent in MDA-MB-453 cell lines. And these selected compounds 6i was led to morphological changes after treatment with MDA-MB-453 cell lines and found clear destabilization of mitochondrial membrane potential behind the observed anticancer activity. This strategy is operationally simple and works with a diverse range of substrates and warrants future investigations for further anticancer drug development.

Abstract 202: Early Reprogramming Derived Cocktail FIJs Supports Cardiomyocyte Proliferation for Heart Regeneration

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Yuan-Yuan Cheng
Keyword(s):  
Morphological Changes ◽  
Heart Function ◽  
Sterol Synthesis ◽  
Heart Regeneration ◽  
Ki 67 ◽  
Adult Mice ◽  
Sequence Of Events ◽  
Proliferative Ability

Although remnant cardiomyocytes (CMs) possess a certain degree of proliferative ability, their regenerative efficiency is too low for cardiac regeneration after injury. As terminally differentiated cells, CMs are considered difficult to successfully reprogram into iPSCs. In this study, a modified reprogramming method was employed to avoid damage to the CMs, and the resulting CM-derived iPSCs were characterized their pluripotency and differentiative abilities. During the early stages of reprogramming, we observed sequential morphological changes in both CMs and non-CMs, but alkaline phosphatase assay revealed that CMs demonstrate a time-delayed sequence of events compared to non-CMs. In order to identify the specific events preceding colony formation, total RNA was purified for microarray analysis on day 0, 2, 4, and 6 of the reprogramming process. There were several interesting changes revealing such as up-regulation of sterol synthesis in non-CMs and down-regulation of chemokines in CMs on day 2. Most importantly, we detected significant enhanced mitosis for CMs on day 2. Nevertheless, typical iPSC-like colonies were clearly observed after 6 days of reprogramming in both cell types. In the purpose of bringing CM back to regain proliferative ability for heart regeneration, the candidates were selected from the microarray results on day 2 of the reprogramming. Triple combined gene cocktail FIJs was determined supporting CM proliferation with 7 times higher Ki-67 + or H3P + population % in neonatal murine CMs in vitro . The same supportive role of FIJs was also confirmed in adult mice showing higher H3P + adult CMs in vivo . Furthermore, heart function detected by echocardiography was significantly improved and less fibrosis shown by trichrome staining after the triple gene cocktail treatment in adult mice suffering from myocardial infarction, and this improvement was owing to the enhanced CM proliferation. In conclusion, triple gene cocktail selected from CM reprogramming day 2 provides valuable information for inducing remnant CM proliferation for heart regeneration.

scholarly journals Soya phytonutrients act on a panel of genes implicated with BRCA1and BRCA2 oncosuppressors in human breast cell lines

2006 ◽  
Vol 95 (2) ◽  
pp. 406-413 ◽  
Author(s):  
Bertrand Caëtano ◽  
Ludovic Le Corre ◽  
Nassera Chalabi ◽  
Laetitia Delort ◽  
Yves-Jean Bignon ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
Response To Treatment ◽  
Breast Cancer Cell Lines ◽  
Human Breast ◽  
Ki 67 ◽  
Breast Cell Lines ◽  
Human Breast Cell ◽  
Mcf 7

Breast cancer is the most common cancer in women and a significant cause of death. Mutations of the oncosuppressor genes BRCA1 and BRCA2 are associated with a hereditary risk of breast cancer, and dysregulation of their expression has been observed in sporadic cases. Soya isoflavones have been shown to inhibit breast cancer in studies in vitro, but associations between the consumption of isoflavone-containing foods and breast cancer risk have varied in epidemiological studies. Soya is a unique source of the phytoestrogens daidzein (4′,7-dihydroxyisoflavone) and genistein (4′,5,7-trihydroxyisoflavone), two molecules that are able to inhibit the proliferation of human breast cancer cells in vitro. The aim of the present study was to determine the effects of genistein (5μg/ml) and daidzein (20μg/ml) on transcription in three human breast cell lines (one dystrophic, MCF10a, and two malignant, MCF-7 and MDA-MB-231) after 72h treatment. The different genes involved in the BRCA1 and BRCA2 pathways (GADD45A, BARD1, JUN, BAX, RB1, ERα, ERβ, BAP1, TNFα, p53, p21Waf1/Cip1, p300, RAD51, pS2, Ki-67) were quantified by real-time quantitative RT-PCR, using the TaqMan method and an ABI Prism 7700 Sequence Detector (Applied Biosystems). We observed that, in response to treatment, many of these genes were overexpressed in the breast cancer cell lines (MCF-7 and MDA-MB-231) but not in the dystrophic cell line (MCF10a).

scholarly journals Development of Functionalized silver Nanoparticles from Allamanda neriifolia Hook and their In Vitro Cytotoxic Effect on MCF-7 Cells

YMER Digital ◽  
2021 ◽  
Vol 20 (12) ◽  
pp. 385-396
Author(s):  
Sumathi R ◽  
◽  
Sivagamasundari K ◽  
Keyword(s):  
Breast Cancer ◽  
Silver Nanoparticles ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Morphological Changes ◽  
Total Phenolic ◽  
Biosynthesized Agnps ◽  
Average Size ◽  
Mcf 7

The present work demonstrates the influence of plant extract composition (antioxidant and total phenolic content) on the size and morphology of the produced AgNPs. In this study, silver nanoparticles (AgNPs) were synthesized using aqueous flower extract of Allamanda neriifolia plant. The biosynthetic procedure was rapid and simple and was easily monitored via colour changes and examined AgNPs (AN-AgNPs) by ultraviolet-visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscope (SEM). The results obtained from various characterizations revealed that average size of synthesized AgNPs was 50 nm and in spherical structure. The anticancer potential of AN-AgNPs was investigated against human breast cancer cells (MCF-7). The cytotoxic response was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), and morphological changes by apoptosis. The biosynthesized AgNPs-induced cell death in MCF- 7 cells suggested the anticancer potential of AN-AgNPs. Therefore, they may be used to treat the breast cancer cells.

scholarly journals High CD45 expression of CD8+ and CD4+ T cells correlates with the size of HIV-1 reservoir in blood

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Stefan Petkov ◽  
Yonas Bekele ◽  
Tadepally Lakshmikanth ◽  
Bo Hejdeman ◽  
Maurizio Zazzi ◽  
...  
Keyword(s):  
T Cells ◽  
Cd4 T Cells ◽  
Tyrosine Phosphatase ◽  
Mass Cytometry ◽  
Tcr Signaling ◽  
Ki 67 ◽  
Proliferation Markers ◽  
Hla Dr ◽  
Hiv 1

AbstractUsing mass cytometry, we investigated the expression of 28 markers on CD8+ and CD4+ T cells from HIV-1 infected patients with a variable size of HIV-1 reservoir defined as high (HR) and low (LR) reservoir; we aimed at identifying phenotypic associations of T cells with size of HIV-1 reservoir. We showed that the frequency of CD45+ CD8+ and CD4+ T cells was directly proportional to the size of HIV-1 reservoir; HR patients had a significantly larger frequency of blood CD45high T cells and higher CD45 expression on both CD8+ and CD4+ T cells. CD45 is a receptor-type protein tyrosine phosphatase essential in TCR signaling. Functional and phenotypical analysis of CD45high cells revealed that they express activation and proliferation markers (CD38 + HLA-DR + and Ki-67) and produce cytokines upon in vitro activation. CD45high T cells also expressed high levels of immune check-point PD-1. Our results link CD45 expression on T cells to HIV-1 reservoir; PD-1 expression on CD45high T cells may contribute to their exhaustion.

scholarly journals IN VITRO EVALUATION OF ANTICANCER POTENTIAL OF ERYTHRINA VARIEGATA L. ON BREAST CANCER CELL LINES

2017 ◽  
Vol 10 (7) ◽  
pp. 305
Author(s):  
Vaishali Rai M ◽  
Vinitha Ramanath Pai ◽  
Samuel Kevin ◽  
Herga P Kedilaya
Keyword(s):  
Cell Lines ◽  
Anticancer Agents ◽  
Morphological Changes ◽  
Treated Group ◽  
Breast Cancer Cell Lines ◽  
Liquid Chromatography Mass Spectrometry ◽  
Erythrina Variegata ◽  
Ic50 Value ◽  
Mcf 7

Objective: Species of Erythrina variegata L. is reported to be used in the treatment of cancer in traditional/folklore medicine which could be explored for their anticancer potential. We aimed to evaluate the anticancer activity of crude extracts of the leaves of E. variegata with two solvents; explore the mechanism of cytotoxicity with the effective extract and correlation with the phytochemicals in the extract.Methods: The extracts with Erythrina variegata L methanol (EVM) and chloroform (EVC) as solvents were screened for cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assay on MDA-MB-231 and MCF-7 cell lines. The effective extract was further evaluated on MDA-MB-231 cells by nucleoprotein content estimation, and cell morphology was studied. High resolution liquid chromatography mass spectrometry (HRLCMS) of EVM was done to find the phytochemical composition.Results: Among the two extracts, EVM was effective at an inhibitory concentration (IC50) value of 92 μg/ml and 143 μg/ml on MCF-7 and MDA-MB-231 cells, respectively. At the IC50 value (143 μg/ml) the nucleoprotein content of the cells was 58.2%, and the apoptotic index was calculated to be 51.8%. EVM treated group showed significant morphological changes suggestive of apoptosis. HRLCMS revealed the presence of rutin, podocarpatriene, and cepharanthine which are known to be cytotoxic.Conclusion: This report is a contribution toward the validation of E. variegata as a potential source of anticancer agents.

scholarly journals MODIFYING EFFECTS OF LACTOFERRIN IN VITRO ON MOLECULAR PHENOTYPE OF HUMAN BREAST CANCER CELLS

2015 ◽  
Vol 37 (3) ◽  
pp. 181-186 ◽  
Author(s):  
V F Chekhun ◽  
I V Zalutskii ◽  
L A Naleskina ◽  
N Yu Lukianova ◽  
T M Yalovenko ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Lines ◽  
Human Breast Cancer ◽  
Resistant Cell ◽  
Molecular Profile ◽  
Human Breast ◽  
Molecular Phenotype ◽  
Ki 67 ◽  
Mcf 7

Aim: To assess the role of endogenous lactoferrin (LF) in the formation of the molecular phenotype of human breast cancer (BC) cell lines with varying degrees of malignancy, including cisplatin/doxorubicin resistant cell lines, and identify possible impact of exogenous LF. Materials and Methods: 5 breast cell lines of different origin — MCF-10 A, MCF-7, including doxorubicin/ cisplatin resistant ones, T47D, MDA-MB-231, and MDA-MB-468. Immunocytochemistry: e xpression of LF, Ki-67, adhesion molecules E- and N-cadherin, CD44, CD24 rating the invasive potential of cells. Results: Expression of LF in human BC cell lines varies. It is associated with the heterogeneity of molecular profiles of cell lines in terms of adhesion. A link has been established between the level of LF expression in the resistant cell line MCF-7/CP and MCF-7/Dox, features of their molecular profile and invasive properties. Exogenous LF was shown to be capable of modifying the molecular profile and invasive properties of all the studied cell lines including resistant ones (MCF-7/CP and MCF-7/Dox). Conclusions: The sensitivity of cytostatic-resistant cell lines (MCF-7/CP and MCF-7/Dox) tends to increase under the influence of exogenous LF. It is likely that this effect is due to LF-mediated inhibition of the expression of proteins associated with drug resistance.

scholarly journals In Vitro Cytotoxicity of Mesoporous SiO2@Eu(OH)3Core-Shell Nanospheres in MCF-7

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
M. Atif ◽  
Muhammad Fakhar-e-Alam ◽  
Najeeb Abbas ◽  
Maqsood A. Siddiqui ◽  
Anees A. Ansari ◽  
...  
Keyword(s):  
Morphological Analysis ◽  
Morphological Changes ◽  
Sol Gel ◽  
Average Diameter ◽  
In Vitro Cytotoxicity ◽  
Core Shell ◽  
Clinical Approach ◽  
Transmission Electron ◽  
Mcf 7

Initially, the sample was synthesized by a modified sol-gel process. Morphological analysis of growth SiO2@Eu(OH)3was confirmed by applying field emission transmission electron microscopy (high and low resolution FETEM). The images confirmed the average diameter of mesoporous SiO2@Eu(OH)3core-shell nanospheres (~392–400 nm) with a silica core of ~230 nm in diameter and a shell composed of europium hydroxide ~162 nm (thickness). Moreover, an absorption band at 280 nm was confirmed which initiates from the europium hydroxide. The photoluminescence spectrum of the nanosphere was also recorded at ambient temperature under the excitation of 3.82 eV. Cytotoxic studies in vitro were performed by applying MTT, NR assays, and morphological analysis. Morphological changes and % loss in cellular viability was assessed in human breast cancer cells (MCF-7) labeled with mesoporous SiO2@Eu(OH)3core-shell nanospheres at different concentrations ranging from 10 µg/mL to 200 µg/mL. Current study demonstrates the quite rational strategy which might be useful in future clinical approach/applications.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

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