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Assessment of Airway Smooth Muscle Growth and Division: In Vitro Studies
Published in Alastair G. Stewart, AIRWAY WALL REMODELLING in ASTHMA, 2020
DNA synthesis measured by flow cytometry is based upon the principle that cells in different stages of the cell cycle have different but predictable amounts of DNA.64,65 By the use of fluorochromes (e.g., propidium iodide, ethidium bromide, and acridine orange) as nuclear dyes which stoichiometrically bind to DNA, it is possible to distinguish in a single parameter three compartments of the cell cycle based solely on DNA content of exponentially dividing cells. These are the G0/G1, phase with diploid amounts of DNA, G2 and mitosis with tetraploid quantities of DNA, and S phase with variable intermediate amounts of DNA. A typical DNA distribution of exponentially growing smooth muscle cells is illustrated in Figure 3. Examples of typical DNA distribution profiles at different phases of the cell cycle are shown in Figure 2. A more detailed account of the application of flow cytometry to cell cycle analysis is described elsewhere.65–67 Clearly, flow cytometry is a very powerful tool which combines detection of DNA synthesis and immunocytochemical markers to provide indices of cell counting, ploidy, and the DNA content in cells.
Cytotoxic Phenanthridone Alkaloid Constituents of the Amaryllidaceae
Published in Spyridon E. Kintzios, Maria G. Barberaki, Evangelia A. Flampouri, Plants That Fight Cancer, 2019
Jerald J. Nair, Johannes van Staden
Pancratistatin together with two of its ester analogs (20, 22) at concentrations less than 1 µM exhibited strong cytostatic activity in 3Y1 rat embryo fibroblasts (Mutsuga et al. 2002). Cell cycle analysis showed that when cells were arrested at the G0/G1 phase via serum deprivation, progression to the S phase was hindered by all three compounds (Mutsuga et al. 2002). Furthermore, cells synchronized at the late G1/early S phase by hydroxyurea treatment were blocked in progressing through the S phase by compound 20, whilst compound 22 and pancratistatin did not affect cell cycle progress but in fact retarded it (Mutsuga et al. 2002). When the effects of 20 and 22 were evaluated in promyelocytic HL-60RG leukemia cells synchronized at the G0/G1 phase, the cells accumulated in the sub G0/G1 phase without progress to the S phase, indicative of apoptotic cells (Mutsuga et al. 2002).
Site-Selective cAMP Analogs in the Arrest of Cancer Cell Growth
Published in Robert I. Glazer, Developments in Cancer Chemotherapy, 2019
The cell cycle analysis was performed to examine whether the reduced cell proliferation observed in the cancer cell lines after treatment with the analogs was due to a specific block in one phase of the cell cycle. It was found that the fractions of breast and colon cancer cells in G1, S, and G2/M phases were not appreciably different between the control cells (untreated) and the cells treated with the analogs. Thus, the inhibition of cell growth induced by the cAMP analogs was not associated with a specific block in one phase of the cell cycle, confirming a previous report41 on cAMP-induced inhibition of human breast cancer cell lines. In contrast, 8-Cl-adenosine-treated cells exhibited a G1 block of cell cycle, indicating that the growth-inhibitory effect of 8-Cl-cAMP was not due to the cytotoxic effects of its adenosine metabolite.
Dorycnium pentaphyllum Extract Has Antiproliferative Effect on Human Cervix and Colon Cancer Cells
Published in Nutrition and Cancer, 2020
Selim Demir, Serap Ozer Yaman, Sila Ozlem Sener, Elif Ayazoglu Demir, Rezzan Aliyazicioglu, Ufuk Ozgen, Ahmet Mentese, Orhan Deger, Yuksel Aliyazicioglu
Cell cycle analysis was performed using a commercial kit according to the manufacturer's recommendations. HeLa cells were treated with 25–100 μg/mL concentrations of D. pentaphyllum extract for 72-h, before being harvested, and washed twice with buffer solution. WiDr cells were treated with 42.5–170 μg/mL concentrations of D. pentaphyllum extract for 72 h, before being harvested, and washed twice with buffer solution. Cells were then incubated with trypsin solution for 10 min followed by trypsin inhibitor and RNase solution for 10 min. Finally, the cells were further treated with propidium iodide (PI) staining on ice for 10 min. Data from 30,000 cells per sample were collected and analyzed on a flow cytometer (BD Accuri C6, MI, USA). The percentage of cells in cycle phases was determined using MODFIT 3.0 verity software. The results were finally compared with those of the untreated cells.
Dicranopteris linearis extract inhibits the proliferation of human breast cancer cell line (MDA-MB-231) via induction of S-phase arrest and apoptosis
Published in Pharmaceutical Biology, 2018
Aifaa Akmal Baharuddin, Rushduddin Al Jufri Roosli, Zainul Amiruddin Zakaria, Siti Farah Md. Tohid
To identify whether the growth inhibitory effect of MEDL was caused by specific disruption of the cell cycle-related event, the DNA content of MDA-MB-231 cells were measured using a flow cytometric analysis. Flow cytometry was used to monitor cell cycle distribution in MDA-MB-231 cells after exposure to MEDL at IC50 concentration for 24, 48, and 72 h. Measurements of the cell cycle analysis were conducted by calculating the proportion of cells in each cycle from a total of 10,000 cells in three independent assays. Flow cytometry also enables the identification of cell distribution in diverse phases of cell cycle. There are four distinct phases that could be distinguished in a proliferating cell population: the G1, S (DNA synthesis), G2 and M phase (mitosis) (Nunez 2001).
Boosting curcumin activity against human prostatic cancer PC3 cells by utilizing scorpion venom conjugated phytosomes as promising functionalized nanovesicles
Published in Drug Delivery, 2022
Mohammed W. Al-Rabia, Nabil A. Alhakamy, Waleed Y. Rizg, Adel F. Alghaith, Osama A. A. Ahmed, Usama A. Fahmy
The outcome of cell cycle analysis showed that cells of the control group were present at a maximum in the G0–G1 and S phases with optimum proliferation. The percentages of cells present in the G2-M and pre-G1 phases signify apoptotic activity or cell cycle arrest. Thus, it can be observed from Figure 5 that CUR–PL–SV produced potent anticancer effects against PC3 cell line via cell cycle arrest in the pre-G1 phase compared to the effects produced by CUR and PL-SV individually. However, CUR showed a superior anticancer impact on the G2-M phase compared to CUR–PL–SV. However, overall, CUR–PL–SV showed superior anticancer potential and arrested cell cycle and hence, inhibited proliferation. The percentage of data was estimated after excluding dead cells in the pre-G phase.