Differential radiation effects in smokers – culture time dependence of the yield of gamma ray-induced chromosome damage in first division metaphases

Abstract

Purpose: Telomeric associations (TA) and unstable chromosomal aberration (CA) transmission through M1-M4 metphases (first to fourth division) in γ-ray irradiated G0 lymphocytes in 2 smokers were examined, since TA in conventionally stained chromosomes were reported earlier as a sensitive cytogenetic marker in mutagen-exposed populations. The purpose of the present study is an extension of our earlier studies on unstable CA transmission through successive mitotic divisions.

Materials and methods: The bromodeoxyuridine (BrdU) incorporation and fluorescence plus giemsa (FPG) method for M1-M5 metaphase analysis was carried out at 50, 72, 96 h to analyse TA and CA in conventionally and FPG stained chromosomes after irradiation of human blood samples with 3 Gy of γ-rays. In situ hybridization (ISH) with enzymatic/fluorescence detection was used to analyse radiation-induced aneuploidy and TA. Analysis was carried out on sister chromatid exchanges (SCE) in M2 cells at 72 h and micronuclei (MN) at 24, 50, 72, 96 h.

Results: TA, corroborated by the absence of acentric fragments, were not detected in conventional/FPG stained/ISH chromosomes. Chromosome 21 aneuploidy was observed. Significant differences in mean frequencies of dicentrics/micronuclei (MN)/SCE with high frequency cells (HFC) were found in smokers after irradiation compared to non-smokers. Higher radiation induced CA in M1 cells were found with extended culture time. Induction of giant cells with mirror dicentrics, tricentrics and rings were found.

Conclusion: TA in conventional or FPG stained metaphase chromosomes is not a sensitive cytogenetic marker for mutagen exposed population screening. Higher radiation induced CA frequencies in M1 cells with extended culture time were indicative of a delay in cell cycle progression of aberrant cells or different lymphocyte subset populations. Bridge-breakage-fusion (BBF) events due to dicentrics may be instrumental in the perpetuation of chromosomal instability. Differential effects were noted in radiation-induced dicentric, SCE and MN frequencies in smokers compared to non-smokers. Heavy smoking could be a confounding variable in chromosome-based biodosimetry and biomonitoring studies. Giant cells may denote a switch to amitotic modes of cell survival, providing additional mechanisms of genotoxic resistance.

Keywords:

• Ionising radiation
• chromosome abberations
• FISH
• micronuclei
• sister chromatid
• exchanges
• human lymphocytes