title: Propidium iodide staining method for testing the cytotoxicity of 2,4,6-trichlorophenol and perfluorooctane sulfonate at low concentrations with Vero cells.
abstract: In this article, the feasibility of propidium iodide (PI) staining based on cell membrane damage as a sensitive and quantitative method to test the cytotoxicity of typical lipophilic compounds including 2,4,6-trichlorophenol (TCP) and perfluorooctane sulfonate (PFOS) was examined. The sensitivity of PI staining was compared to that of the methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. We found a good correlation between PI uptake and increasing concentrations of TCP (10-50 μM) and PFOS (20-50 μM) at lower concentrations than those between growth inhibition ratio determined by the MTT assay and increasing concentrations of TCP (150-400 μM) and PFOS (100-500 μM). These findings indicated that the PI staining was more sensitive than the MTT assay. Furthermore, both the amount of PI uptake and the ratio of morphologically changed cells increased with increasing TCP concentrations, suggesting that PI staining may be a suitable quantitative substitute for the original method based on the observation of morphological changes in Vero cells. Our data also suggest that it is possible that the cell membrane damage that resulted in increased PI uptake may be due to variations in the phospholipid and protein content of the membrane, which are affected by interactions between the lipophilic compounds and components of the cell membrane.
authors.affiliation: College of Environmental Science and Engineering, State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai, China.
authors.forename: Ting T
authors.initials: TT
authors.lastname: Liao
authors.forename: Ru W
authors.initials: RW
authors.lastname: Jia
authors.forename: Yan L
authors.initials: YL
authors.lastname: Shi
authors.forename: Jian W
authors.initials: JW
authors.lastname: Jia
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Hong
authors.initials: H
authors.lastname: Chua
mesh_terms: D017738:Alkanesulfonic Acids / Q000633:toxicity*; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D002733:Chlorophenols / Q000633:toxicity*; D004396:Coloring Agents; D004785:Environmental Pollutants / Q000633:toxicity; D005466:Fluorocarbons / Q000633:toxicity*; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011419:Propidium; D013778:Tetrazolium Salts; D013844:Thiazoles; D014709:Vero Cells
chemical_list: D017738:Alkanesulfonic Acids; D002733:Chlorophenols; D004396:Coloring Agents; D004785:Environmental Pollutants; D005466:Fluorocarbons; D008565:Membrane Proteins; D010743:Phospholipids; D013778:Tetrazolium Salts; D013844:Thiazoles; D011419:Propidium; C076994:perfluorooctane sulfonic acid; C022616:thiazolyl blue; C024564:2,4,6-trichlorophenol