In vitro Cytotoxic Effects of Control and Selenium Conjugated Transferrins against Leukemia Cell Lines K562 and THP-1
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2019, Vol 13, Issue 3
Abstract
Human leukemia is a malignancy of hematopoietic stem cells with impaired differentiation and uncontrolled proliferation. Many leukemia cell lines over-express the trans-membrane transferrin receptor (TfR) which endocytoses iron (Fe) bound Holo-transferrin (Holo-Tf). Over-expression of the TfR has permitted transferrin-mediated drug delivery targeting leukemia cells. Selenides (RSe-), can generate free radicals, i.e.; superoxide (O2.-) and other Reactive Oxygen Species (ROS) by redox cycling; oxidizing thiols (RSH) within cells following endocytosis. Superoxide generation by selenium (Se) is reported to cause cell apoptosis/necrosis due to increased endogenous oxidative stress. In the present study, Se as ROCH2CH2SeCN was covalently attached to Lysine residues of Apo-, Holo-Tf, and Human Serum Albumin (HuSA) using a Se-modified Bolton-Hunter reagent. The Se conjugated proteins were used to target the TfR and experimentally treat K562 and THP-1 leukemia cell lines. Cells were also treated with sodium selenite (10 μgSe/well) as a positive control known to cause cell death. Holo-transferrin (Tf) binds iron (Fe) from the epithelial intestine cells and transports it into liver tissue and other cells [1]. Two types of transferrin receptors are found on the cell surface of plasma membranes; transferrin receptor 1 (TfR1) [2] and transferrin receptor 2 (TfR2) [3]. Holo-transferrin has a higher affinity for TfR1 than TfR2 and TfR1 is more abundantly expressed on cell membranes than TfR2. Almost every cell that requires Fe, expresses TfR1 having two distinct cytoplasmic domains; an extended anchored hydrophobic membrane and exocytic region of the protein [4]. Extensive research now suggests that Fe bound transferrin is internally transported by mediated endocytosis [5]. On rapidly dividing cancer cells, approximately 10,000 to 100,000 TfR1 receptor molecules are over-expressed per cell [6]. On normal cells, however, expression of TfR1 is generally low or undetectable. Holo-transferrin delivers Fe to human leukemia hematopoietic stem cells, cells characterized by uncontrolled proliferation.Following selenium conjugation, the proteins were quantified for Se by ICP-MS (μgSe/mg protein). K562 and THP-1 leukemia cell lines under conventional culture conditions were treated in 24-well plates with different concentrations of Se conjugated Apo-, Holo-Tf, and HuSA proteins and selenite at 24, 48, 72, 96, and 120 hours. Cells responded with divisional arrest to Se concentrations with photographic changes in cell morphology in a dose and time dependent manner as determined by Trypan Blue absorption, MTT, and Flow Cytometry assays. Caspase-3 was increased (p < 0.05) in both K562 and THP-1 cells after Se treatments, indicating the arrest of cell proliferation was partially apoptotic.
Authors and Affiliations
Debalina Goswami Sewell, Priyanka Bapat, L Mallory Boylan, Julian E Spallholz
Keywords
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- EP ID EP594442
- DOI 10.26717/BJSTR.2019.13.002402
- Views 134
- Downloads 0
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