Flow cytometric analysis of mitotic cycle perturbation by chemical carcinogens in cultured epithelial cells. [Effects of benzo(a)pyrene-diol-epoxide on mitotic cycle of cultural mouse liver epithelial cells]

Thesis/Dissertation · Tue Aug 01 00:00:00 EDT 1978

DOI:https://doi.org/10.2172/6503301· OSTI ID:6503301

Pearlman, Andrew Leonard ^[1]

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1. Univ. of California, Berkeley, CA (United States)

A system for kinetic analysis of mitotic cycle perturbation by various agents was developed and applied to the study of the mitotic cycle effects and dependency of the chemical carcinogen benzo(a)pyrene-diolepoxide, DE, upon a mouse lever epithelial cell line, NMuLi. The study suggests that the targets of DE action are not confined to DNA alone but may include cytoplasmic structures as well. DE was found to affect cells located in virtually every phase of the mitotic cycle, with cells that were actively synthesizing DNA showing the strongest response. However, the resulting perturbations were not confined to S-phase alone. DE slowed traversal through S-phase by about 40% regardless of the cycle phase of the cells exposed to it, and slowed traversal through G₂M by about 50%. When added to G₁ cells, DE delayed recruitment of apparently quiescent (G₀) cells by 2 hours, and reduced the synchrony of the cohort of cells recruited into active proliferation. The kinetic analysis system consists of four elements: tissue culture methods for propagating and harvesting cell populations; an elutriation centrifugation system for bulk synchronization of cells in various phases of the mitotic cycle; a flow cytometer (FCM), coupled with appropriate staining protocols, to enable rapid analysis of the DNA distribution of any given cell population; and data reduction and analysis methods for extracting information from the DNA histograms produced by the FCM. The elements of the system are discussed. A mathematical analysis of DNA histograms obtained by FCM is presented. The analysis leads to the detailed implementation of a new modeling approach. The new modeling approach is applied to the estimation of cell cycle kinetic parameters from time series of DNA histograms, and methods for the reduction and interpretation of such series are suggested.

• Research Organization: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
• Sponsoring Organization: USDOE
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 6503301
• Report Number(s): LBL-8169
• Resource Relation: Other Information: Thesis
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
CELL CYCLE
MITOTIC DELAY
CARCINOGENS
BIOLOGICAL EFFECTS
INHIBITION
CELL FLOW SYSTEMS
PERFORMANCE
BENZOPYRENE
CELL CULTURES
DATA ANALYSIS
DATA PROCESSING
EPITHELIUM
EPOXIDES
GLYCOLS
LIVER
MATHEMATICAL MODELS
MICE
ALCOHOLS
ANIMALS
AROMATICS
BODY
CONDENSED AROMATICS
DIGESTIVE SYSTEM
GLANDS
HYDROCARBONS
HYDROXY COMPOUNDS
MAMMALS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
ORGANS
PROCESSING
RODENTS
TISSUES
VERTEBRATES
560301* - Chemicals Metabolism & Toxicology- Cells- (-1987)
550300 - Cytology