New Methods for Cell Cycle Analysis

Fang, Hanshu; Lang, Ming-Fei; Sun, Jing

doi:10.1016/s1872-2040(19)61186-2

Cited by 10 publications

(6 citation statements)

References 40 publications

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“…Further experiments were realized to assess the potential mechanisms responsible for the cytotoxic effects of DOX combined with hyperthermia. Cell cycle is a cellular process common to all cells and important for cell proliferation [ 47 ]. To avoid errors, this process is controlled by specific molecular mechanisms that guarantee genome integrity [ 48 ].…”

Section: Discussionmentioning

confidence: 99%

Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

Salvador

Bastos

Oliveira

2021

IJMS

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Melanoma is the deadliest form of skin cancer, and its incidence has alarmingly increased in the last few decades, creating a need for novel treatment approaches. Thus, we evaluated the combinatorial effect of doxorubicin (DOX) and hyperthermia on A375 and MNT-1 human melanoma cell lines. Cells were treated with DOX for 24, 48, and 72 h and their viabilities were assessed. The effect of DOX IC10 and IC20 (combined at 43 °C for 30, 60, and 120 min) on cell viability was further analyzed. Interference on cell cycle dynamics, reactive oxygen species (ROS) production, and apoptosis upon treatment (with 30 min at 43 °C and DOX at the IC20 for 48 h) were analyzed by flow cytometry. Combined treatment significantly decreased cell viability, but not in all tested conditions, suggesting that the effect depends on the drug concentration and heat treatment duration. Combined treatment also mediated a G2/M phase arrest in both cell lines, as well as increasing ROS levels. Additionally, it induced early apoptosis in MNT-1 cells, while in A375 cells this effect was similar to the one caused by hyperthermia alone. These findings demonstrate that hyperthermia enhances DOX effect through cell cycle arrest, oxidative stress, and apoptotic cell death.

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Section: Discussionmentioning

confidence: 99%

Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

Salvador

Bastos

Oliveira

2021

IJMS

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“…Immunohistochemistry-based cell cycle detection (iCCD) 53,54 • Geminin: Highest expression in S/G2/M phase and degrades after M phase • Chromatin licensing and DNA replication factor 1 (Cdt1): Highest expression in G1 phase and degrades before S phase • Gamma H2A.X: Marker for DNA double-strand breaks (DSBs) 55 Fluorescence in situ hybridization (FISH) 56 • Ploidy assessment by centromere and α-satellite DNA assessment probes Fluorescent ubiquitination-based cell cycle indicator (FUCCI) 57 • Fluorescence-labelled proteins tagged with Cdt 1 and Geminin • Time-lapse photographs are used to identify both temporality and spatial organization of cells in culture according to their stage in cell cycle Radioisotope-labeled DNA 58 • Incorporating radioisotope (now replaced by nucleoside analogues) into the DNA to calculate the time spent by a cell across the cell cycle stages Flow cytometry 2,3,8,9,52 Univariate • Using DNA binding dyes that enable only cellular DNA content assessment • Applicable in samples that have only/predominantly tumor cells…”

Section: Technique Brief Descriptionmentioning

confidence: 99%

Flow Cytometric Ploidy Analysis in Acute Lymphoblastic Leukemia and Plasma Cell Myeloma

Bommannan

2023

Indian J Med Paediatr Oncol

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Identification of underlying cytogenetic (CG) aberrancies plays a significant role in risk stratification of hematological malignancies. These abnormalities can be due to aberrancies that affect the number or structure of chromosomes. Numerical chromosomal abnormalities are called aneuploidies, which result from either gain or loss of whole chromosomes. Ploidy assessment by CG is a laborious and less sensitive technique. With the aid of fluorescent nucleic acid binding dyes, the total DNA content and different phases of the cell cycle specific to any population of interest can be deciphered and analyzed by flow cytometry (FCM). DNA index (DI), a parameter derived by FCM DNA analysis, is equivalent to conventional CG-based ploidy assessment. In this study, the technical aspects and implications of FCM DNA assessment among patients diagnosed with acute lymphoblastic leukemia and plasma cell myeloma are discussed.

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“…For example, the use of very common synchronization protocols based on thymidine, mimosine, or aphidicolin may lead to growth imbalance and can also induce imbalance in the expression of cell cycle regulatory proteins such as cyclins B1, A, and E [ 3 ]. Less common and emerging techniques of cell cycle analysis are reviewed, e.g., in [ 4 ] or [ 5 ].…”

Section: Introductionmentioning

confidence: 99%

Basic Methods of Cell Cycle Analysis

Ligasová

Frydrych

Koberna

2023

IJMS

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Cellular growth and the preparation of cells for division between two successive cell divisions is called the cell cycle. The cell cycle is divided into several phases; the length of these particular cell cycle phases is an important characteristic of cell life. The progression of cells through these phases is a highly orchestrated process governed by endogenous and exogenous factors. For the elucidation of the role of these factors, including pathological aspects, various methods have been developed. Among these methods, those focused on the analysis of the duration of distinct cell cycle phases play important role. The main aim of this review is to guide the readers through the basic methods of the determination of cell cycle phases and estimation of their length, with a focus on the effectiveness and reproducibility of the described methods.

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New Methods for Cell Cycle Analysis

Cited by 10 publications

References 40 publications

Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

Hyperthermia Enhances Doxorubicin Therapeutic Efficacy against A375 and MNT-1 Melanoma Cells

Flow Cytometric Ploidy Analysis in Acute Lymphoblastic Leukemia and Plasma Cell Myeloma

Basic Methods of Cell Cycle Analysis

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