Flow Cytometric Analysis of Fluorescent Estrogen Binding in Cancer Cell Suspensions

2019 ◽  
pp. 95-110
Author(s):  
Chris Benz ◽  
Israel Wiznitzer ◽  
Sin Hang Lee
Keyword(s):  
Cancer Cell ◽  
Flow Cytometric Analysis ◽  
Cell Suspensions ◽  
Flow Cytometric ◽  
Estrogen Binding

Evaluation on the IGF-IR Inhibitor CP-751,871, alone and in combination with paclitaxel in lung and colon cell lines

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e22125-e22125 ◽  
Author(s):  
R. Garcia-Carbonero ◽  
M. T. Agullo-Ortuño ◽  
F. Lopez-Rios ◽  
C. V. Diaz-Garcia ◽  
A. Cortijo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Colon Cancer ◽  
Mrna Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Flow Cytometric Analysis ◽  
Colon Cancer Cell ◽  
Sequential Treatment ◽  
Flow Cytometric ◽  
Colon Cancer Cell Lines

e22125 Background: Signaling through the insulin-like growth factor 1 receptor (IGF-IR) has been implicated in the resistance to a number of clinically relevant anti-cancer agents. The aim of this study was to assess the direct anti-tumour effects of CP-751,871, alone and in combination with paclitaxel in lung and colon cancer cell lines. Methods: Four lung and four colon cancer cell line where treated with the IGF-IR inhibitor CP-751,871 and/or Paclitaxel in simultaneous or sequential treatments. Response to treatments was evaluated by WST-1 cell survival assays. Flow cytometric analysis was used to estimate the effect on the cell cycle and apoptosis. IGF-IR mRNA expression was determined by quantitative real-time PCR and relative gene expression values were calculated by the ΔΔCt method. Results: No correlation between basal IGF-IR mRNA expression and CP-751,871 response was observed in cell lines tested. Flow cytometric analysis demonstrated that CP-751,871 enhanced cell cycle arrest at the G1/G0 checkpoint with minimal effects on apoptosis. Combined simultaneous and, more strongly, sequential treatment with CP- 751,871 and Paclitaxel showed improved response in cell growth inhibition on HCC78, H1299, Colo205 and HT29 cell lines, and was statistically superior to Paclitaxel alone (p< 0.001) and CP-751,871 alone (p< 0.001). In H460, LS180 and DLD-1 cell lines, concomitant, but no sequential treatment resulted in antagonistic interactions. Conclusions: CP-751,871 showed a modest anti-tumour activity, predominantly cytostatic, against lung and colon cancer cell lines, that is not related to the mRNA expression. CP-751,871 tends to enhance the effects of paclitaxel in vitro, depending on sequence of administration and on the model system used. No significant financial relationships to disclose.

scholarly journals ALTERING THE EXPRESSION OF P-GLYCOPROTEIN RESPONSIBLE FOR CHEMOTHERAPEUTIC DRUG RESISTANCE IN HEPG2 CELL LINE POST-TREATMENT WITH UREA AND β-MERCAPTOETHANOL

2019 ◽  
pp. 284-288
Author(s):  
PROMA CHAKRABORTY ◽  
MANI RAMAKRISHNAN ◽  
KUMAR KM
Keyword(s):  
Cell Line ◽  
Anticancer Drugs ◽  
Cancer Cell ◽  
Mtt Assay ◽  
Hepg2 Cells ◽  
Flow Cytometric Analysis ◽  
Post Treatment ◽  
Lead Compounds ◽  
Flow Cytometric ◽  
P Glycoprotein

Objective: The objective of this study is to alter the expression of p-glycoprotein (p-gp) pump proteins in HepG2 cells after treating with urea and β-mercaptoethanol (BME) (lead compounds). The most common cause for resistance to a broad range of anticancer drugs is influenced by overexpression of p-gp pumps that detect and eject anticancer drugs from the cancer cell. Altering the expression of these proteins will reduce the efflux action and enhance the drug retention eventually killing the cancer cell. Materials and Methods: 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyl tetrazolium bromide (MTT) assay was carried out to measure the cell viability (HepG2 cells) post-treatment with the lead compounds followed by flow cytometric analysis for protein expression studies. Results: MTT assay confirms that the viability of HepG2 cells reduces as the concentrations of the lead compounds are increased. Flow cytometric analysis confirms reduced p-gp expression in HepG2 cells post-treatment with urea and BME. Compare to BME, urea turns out to be a potential compound in altering the expression of p-gp. Conclusion: The present cell line study confirms that urea and BME are potential compounds which are able to reduce the p-gp expression inHepG2 cells.

Multiparametric flow cytometric analysis in a breast cancer cell line (MCF-7)

2002 ◽  
Vol 28 (3) ◽  
pp. 141-148 ◽  
Author(s):  
Gui Se Ra Lee ◽  
Ki Sung Ryu ◽  
Jong Gu Rha ◽  
Soo Pyung Kim ◽  
Sung Eun Namkoong ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Breast Cancer Cell Line ◽  
Flow Cytometric Analysis ◽  
Cancer Cell Line ◽  
Flow Cytometric ◽  
Mcf 7

In situ cell kinetics studies on human neuroectodermal tumors with bromodeoxyuridine labeling

1986 ◽  
Vol 64 (3) ◽  
pp. 453-459 ◽  
Author(s):  
Takao Hoshino ◽  
Tadashi Nagashima ◽  
Judith A. Murovic ◽  
Charles B. Wilson ◽  
Michael S. B. Edwards ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Single Cell ◽  
Flow Cytometric Analysis ◽  
S Phase ◽  
Cell Suspensions ◽  
Phase Fraction ◽  
Flow Cytometric ◽  
Neuroectodermal Tumors

✓ Thirty-eight patients undergoing surgical removal of neuroectodermal tumors of the central nervous system were given a 1-hour intravenous infusion of bromodeoxyuridine (BUdR), 150 to 200 mg/sq m, to label tumor cells in the deoxyribonucleic acid (DNA) synthesis phase (S-phase). The excised tumor specimens were divided into two portions: one was fixed with 70% ethanol and embedded in paraffin and the other was digested with an enzyme cocktail to make a single-cell suspension. The paraffin-embedded tissues were stained by an indirect peroxidase method using anti-BUdR monoclonal antibody (MA) as the first antibody. Single-cell suspensions were reacted with fluorescein isothiocyanate (FITC)-conjugated anti-BUdR MA's for flow cytometric analysis. S-phase cells that had incorporated BUdR into their DNA were well stained by both methods. The percentage of BUdR-labeled cells, or S-phase fraction, was calculated in tissue sections by microscopic examination and in single-cell suspensions by flow cytometric analysis. The biological malignancy of the tumors was reflected in the S-phase fractions, which were 5% to 20% for glioblastoma multiforme, medulloblastoma, and highly anaplastic astrocytoma, but less than 1% in most moderately anaplastic astrocytomas, ependymomas, and mixed gliomas. Two juvenile pilocytic astrocytomas and two low-grade astrocytomas from children had high S-phase fractions despite the fairly benign and slow-growing nature of these tumors. These results indicate that the S-phase fraction obtained immunocytochemically with anti-BUdR MA's may provide useful information in estimating the biological malignancy of human central nervous system tumors in situ.

Apoptotic Effects of Melittin on 4T1 Breast Cancer Cell Line is associated with Up Regulation of Mfn1 and Drp1 mRNA Expression

2020 ◽  
Vol 20 (7) ◽  
pp. 790-799 ◽  
Author(s):  
Farnaz D. Moghaddam ◽  
Pejman Mortazavi ◽  
Somayeh Hamedi ◽  
Mohammad Nabiuni ◽  
Nasim H. Roodbari
Keyword(s):  
Breast Cancer ◽  
Mrna Expression ◽  
Hemolytic Activity ◽  
Breast Cancer Cells ◽  
Flow Cytometric Analysis ◽  
Control Group ◽  
Flow Cytometric ◽  
4T1 Cells ◽  
4T1 Breast Cancer ◽  
Apoptotic Effects

Background and Purpose: Melittin, as the main ingredient of honeybee venom, that has shown anticancer properties. The present study aimed at investigating the cytotoxic impacts of melittin on 4T1 breast cancer cells. Methods: Hemolytic activity of different concentrations (0.125, 0.25, 0.5, 1, 2, 4, 8μg/ml) of melittin was assayed and then cytotoxicity of selected concentrations of melittin (2, 4, 8, 16, 32, and 64μg/ml), 2 and 4μg/ml of cisplatin and 0.513, 0.295 and 0.123μg/ml of doxorubicin was evaluated on 4T1 cells using MTT assay. We used Morphological evaluation and flow cytometric analysis was used. Real time PCR was also used to determine mRNA expression of Mfn1 and Drp1 genes. Results: All compounds showed anti-proliferative effects on the tumor cell line with different potencies. Melittin had higher cytotoxicity against 4T1 breast cancer cells (IC50= 32μg/ml-72h) and higher hemolytic activity (HD50= 1μg/ml), as compared to cisplatin and doxorubicin. Mellitin at 16 and 32μg/ml showed apoptotic effects on 4T1 cells according to the flow cytometric analysis. The Real time PCR analysis of Drp1 and Mfn1 expression in cells treated with 16μg/ml of melittin revealed an up-regulation in Drp1 and Mfn1 genes mRNA expression in comparison with control group. Treatment with 32μg/ml of melittin was also associated with a rise in mRNA expression of Drp1 and Mfn1 as compared to the control group. Conclusion: The results of this study showed that melittin has anticancer effects on 4T1 cell lines in a dose and time dependent manner and can be a good candidate for further research on breast cancer treatment.

Sign in / Sign up

Export Citation Format

Share Document