title: Inhibition of gap junctional intercellular communication in heptachlor- and heptachlor epoxide-treated normal human breast epithelial cells.
abstract: Based on the concern of organochlorides in the environment and in human tissue, this study was designed to determine whether various noncytotoxic levels of heptachlor and heptachlor epoxide could inhibit, reversibly, gap junctional intercellular communication in human breast epithelial cells (HBEC). Cytotoxicity and gap junctional intercellular communication (GJIC) were evaluated by lactate dehydrogenase assay and fluorescence redistribution after photobleaching analysis, respectively. Both heptachlor and heptachlor epoxide were noncytotoxic up to 10 microg/ml. At this concentration, heptachlor and heptachlor epoxide inhibited GJIC of normal human breast epithelial cells after 1 h treatment. Within a 24 h treatment with heptachlor and heptachlor epoxide at 10 microg/ml, recovery of GJIC had not returned. GJIC completely recovered after a 12 h treatment of 1 microg/ml heptachlor epoxide, but it did not recover after a 24 h treatment of 1 microg/ml heptachlor. RT-PCR and Western blots were analyzed to determine whether the heptachlor or heptachlor epoxide might have altered the steady-state levels of gap junction mRNA and/or connexin protein levels or phosphorylation state. No significant difference in the level of connexin 43 (Cx43) message between control and heptachlor-treated cells was observed. Western blot analyses showed hypophosphorylation patterns in cells treated with 10 microg/ml heptachlor and heptachlor epoxide for 1 h with no recovery within 24 h. Immunostaining of Cx43 protein in normal HBEC indicated that heptachlor and heptachlor epoxide caused a loss of Cx43 from the cell membranes at noncytotoxic dose levels. Taken together, these results suggest that heptachlor and heptachlor epoxide can alter GJIC at the post-translational level, and that, under the conditions of exceeding a threshold concentration in the breast tissue containing 'initiated' cells for a long time and not being counteracted by anti-tumor-promoting chemicals, they could act as breast tumor promoters.
authors: Department of Pediatrics/Human Development and Institute of Environmental Toxicology, Michigan State University, East Lansing 48824, USA.
authors: K
authors: K
authors: Nomata
authors: K S
authors: KS
authors: Kang
authors: T
authors: T
authors: Hayashi
authors: D
authors: D
authors: Matesic
authors: L
authors: L
authors: Lockwood
authors: C C
authors: CC
authors: Chang
authors: J E
authors: JE
authors: Trosko
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D001940:Breast / Q000166:cytology*; D002450:Cell Communication / Q000187:drug effects*; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D017630:Connexins / Q000235:genetics / Q000378:metabolism; D003603:Cytotoxins / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000648:ultrastructure; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D017629:Gap Junctions / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics; D006533:Heptachlor / Q000494:pharmacology* / Q000633:toxicity; D006534:Heptachlor Epoxide / Q000494:pharmacology* / Q000633:toxicity; D006801:Humans; D007306:Insecticides / Q000494:pharmacology* / Q000633:toxicity; D008969:Molecular Sequence Data; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D017630:Connexins; D003603:Cytotoxins; D007306:Insecticides; D012333:RNA, Messenger; D006534:Heptachlor Epoxide; D006533:Heptachlor