Abstract
Apoptosis arises from the active initiation and propagation of a series of highly orchestrated specific biochemical events leading to the demise of the cell. It is a normal physiological process, which occurs during embryonic development as well as in the maintenance of tissue homeostasis. Diverse groups of molecules are involved in the apoptosis pathway and it functions as a mechanism to eliminate unwanted or irreparably damaged cells. However, inappropriate induction of apoptosis by environmental agents has broad ranging pathologic implications and has been associated with several diseases including cancer. The toxicity of several heavy metals such as mercury has been attributed to their high affinity to sulfhydryl groups of proteins and enzymes, and their ability to disrupt cell cycle progression and/or apoptosis in various tissues. The aim of this study was to assess the potential for mercury to induce early and late-stage apoptosis in human liver carcinoma (HepG2) cells. The Annexin-V and Caspase 3 assays were performed by flow cytometric analysis to determine the extent of phosphatidylserine externalization and Caspase 3 activation in mercury-treated HepG2 cells. Cells were exposed to mercury for 10 and 48 hours respectively at doses of 0, 1, 2, and 3 microg/mL based on previous cytotoxicity results in our laboratory indicating an LD50 of 3.5 +/- 0.6 microg/mL for mercury in HepG2 cells. The study data indicated a dose response relationship between mercury exposure and the degree of early and late-stage apoptosis in HepG2 cells. The percentages of cells undergoing early apoptosis were 0.03 +/- 0.03%, 5.19 +/- 0.04%, 6.36 +/- 0.04%, and 8.84 +/- 0.02% for 0, 1, 2, and 3 microg/mL of mercury respectively, indicating a gradual increase in apoptotic cells with increasing doses of mercury. The percentages of Caspase 3 positive cells undergoing late apoptosis were 3.58 +/- 0.03%, 17.06 +/- 0.05%, 23.32 +/- 0.03%, and 34.51 +/- 0.01% for 0, 1, 2, and 3 microg/mL of mercury respectively, also indicating a gradual increase in Caspase positive cells with increasing doses of mercury.
Highlights
All forms of mercury are toxic and exert their effects in a number of organs, tissues, and cell systems [1]
Mercury compounds found in the marine environment pose risk to human health through the consumption of contaminated seafood
The content of each vial was transferred to a 75 cm2 tissue culture flask, diluted with DMEM, supplemented with 10% fetal bovine serum (FBS), 1% streptomycin and penicillin, and incubated for 24 hours at 37oC in a 5% CO2 incubator to allow the cells to grow, and form a monolayer in the flask
Summary
All forms of mercury are toxic and exert their effects in a number of organs, tissues, and cell systems [1]. Mercury and its compounds both organic and inorganic are released to the environment as a result of a variety of human activities. It tends to accumulate in the soils and sediments [3]. Excessive levels of mercury in the marine environment can affect marine biota and pose risk to human consumers of seafood. Mercury compounds found in the marine environment pose risk to human health through the consumption of contaminated seafood. It is desirable to minimize such exposure to levels that do not cause adverse effects [4]
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