The effects of twelve chlorinated aliphatic hydrocarbons upon the cell cycle of Chinese hamster cell lines : V79d-MZ and genetically engineered cell lines expressing CYP2E1

• Main Author: Kim, A.
• Published: Swansea University 1998
• Subjects: 577.27
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.637798

Chlorinated organic chemicals are widely used in industry and are present in the environment. Twelve chlorinated aliphatic hydrocarbons, namely carbon tetrachloride, 1,2-dichloroethane, 1,1,1-trichloroethane, 1,1,2-trichloroethane, hexachloroethane, 1,2-dichloroethylene, trichloroethylene, 1,3-dichloropropane, 1,2,3-trichloropropane, 2,3-dichlorobutane, 1-chlorohexane and 1,1,3-trichloropropene were investigated to determine their influence upon the fidelity of cell division in cultured mammalian cells. In order to determine the influence of these chemical compounds upon the fidelity of cell division, a technique known as differential staining of chromosomes and spindle was performed with two genetically engineered cell lines and their parental cell line. The genetically engineered cell lines used in this study expressed a human P450 enzyme, CYP2E1. Four chemicals, trichloroethylene, 1,2-dichloroethane, 2,3-dichlorobutane and 1-chlorohexane required bioactivation in order to induce spindle damage in cultured mammalian cells whereas all the other chemicals were direct-acting spindle poisons with the exception of 1,1,1-trichloroethane which hardly influenced the fidelity of cell division in all three cell lines. Micronucleus assays were performed to study at which one of the four phases in V79d-MZ cell cycle, the cells were most vulnerable to the effects of two selected chlorinated hydrocarbons. To carry out this assay the cells were synchronized using 1-2% of DMSO. The same assay was also used to investigate at which phase of the cell cycle, the enzyme CYP2E1 activates trichloroethylene. The phase at which the V79d-MZ cells were most vulnerable to the chemical effects was mitosis (M). Finally an apoptosis assay was used to investigate whether the chlorinated aliphatic hydrocarbons are inducing cell death: apoptosis or necrosis using annexin-V and from this it was found that 1,2-dichloroethylene induced apoptosis.