title: A correlation between DNA-nuclear matrix binding and relative radiosensitivity in two human squamous cell carcinoma cell lines.
abstract: Three aspects of DNA topology were examined in two human squamous cell carcinoma lines of differing radiosensitivity (SQ-9G, D0 = 1.46 Gy; and SQ-20B, D0 = 2.36 Gy). High-salt-extracted nuclei (nucleoids) were taken from gamma-irradiated cells, stained with ethidium bromide and examined by flow cytometry. After 5 Gy, nucleoids from SQ-9G cells became 30% less efficient at adopting positive DNA supercoils than were unirradiated controls. In contrast, only a 4% difference was found with the radioresistant SQ-20B line. Both lines produced positive supercoils more efficiently after irradiation if first exposed to the topoisomerase II inhibitor VP16. Ethidium bromide titration of nucleoids was consistent with each containing similar numbers and sizes of DNA loops. In each line approximately 30-35% of DNA was accessible to trioxsalen, as shown by inter-strand crosslinking after UV photo-activation. Exhaustive digestion of nuclear DNA by DNase I removed more DNA from the radiosensitive than from the radioresistant cell line (12% vs 28% remaining). This difference was thought to be due to the increased accessibility of SQ-9G DNA in vitro. We suggest that a looser association of SQ-9G DNA with the nuclear matrix both promotes DNase I digestion and affects the ability of SQ-9G nucleoids to maintain positive DNA supercoils after irradiation. These data implicate the DNA matrix attachment region in the expression of radiation sensitivity in the cell lines studied.
authors.affiliation: Department of Immunology, Medical School, University of Birmingham, UK.
authors.forename: A E
authors.initials: AE
authors.lastname: Milner
authors.forename: D J
authors.initials: DJ
authors.lastname: Gordon
authors.forename: B M
authors.initials: BM
authors.lastname: Turner
authors.forename: A T
authors.initials: AT
authors.lastname: Vaughan
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000503:physiopathology*; D004273:DNA, Neoplasm / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D015530:Nuclear Matrix / Q000378:metabolism*; D011836:Radiation Tolerance / Q000502:physiology*; D014407:Tumor Cells, Cultured / Q000528:radiation effects
chemical_list: D004273:DNA, Neoplasm