The Effect of Honey Bee Venom on CD14 Protein Expression as Monocyte Marker in D-Alpha-Tocopheryl Succinate (Vitamin E)-Treated HL-60 Cells

Azam Yarahmadi; Mohammad Nabiuni; Latifeh  Karimzadeh Bardei; Majid Kabuli

doi:10.18502/pbr.v7i4.9374

Azam Yarahmadi Department of Animal Biology, School of Biological Sciences, Kharazmi University, Tehran, Iran
Mohammad Nabiuni Department of Cell and Molecular Biology, School of Biological Sciences, Kharazmi University, Tehran, Iran.
Latifeh Karimzadeh Bardei Department of Biology, School of Science, University of Tehran, Tehran, Iran
Majid Kabuli Department of Medical Genetics, School of Medicine, Tehran University of Medical Science, Tehran, Iran.

DOI: https://doi.org/10.18502/pbr.v7i4.9374

Keywords: Differentiation, Hematopoietic stem cell, Promyelocytic leukemia, p21 protein

Abstract

Background: The membrane form of a Cluster of Differentiation 14 (CD14) is anchored to the phospholipid bilayer via glycosylphosphatidylinositol anchor. This molecule is expressed on the intrinsic surface of monocytes and neutrophils. This protein is not expressed on the HL60 cell surface. It is believed that the differentiation of HL-60 cells stops in the promyelocytic stage. The differentiation of HL-60 cells with compounds such as vitamin A, D, E results in membrane CD14 expression.

Objectives: The objective of this study was an evaluation of CD14 expression by Honey Bee Venom (HBV) in HL60 cells treated by D-alpha-Tocopheryl Succinate (D-α-TS). Methods: HL-60 cells were cultured in RPMI Media 1640 medium and treated with different concentrations of D-α-TS and HBV. Cellular differentiation was tested by Nitro Blue Tetrazolium (NBT) staining, immunocytochemistry, and flow cytometry. The studied data were analyzed using one-way analysis of variance and InStat 3 software.

Methods: HL-60 cells were cultured in RPMI medium and treated with different concentrations of D-α-TS and HBV. Cellular differentiation was tested by NBT staining, immunocytochemistry, and flow cytometry. The studied data were analyzed using one-way analysis of variance and InStat 3 software.

Results: MTT assay demonstrated that HBV and D-alpha-tocopheryl induce death in HL-60 cell lines in a time and dose-dependent manner. Also, Wright-Giemsa, NBT staining showed morphologically differentiation. Immunocytochemistry and flow cytometry analysis shows that cells treated with 6 µg/mL D-α-TS and 2.5 µg/mL HBV for 5 days significantly increase the expression of CD14 in HL-60 compared to cells treated with D-α-TS.

Conclusion: HBV can induce cell death and inhibit cell proliferation. Also, increase differentiation potency of D-α-TS via HBV can increase the differentiation by inhibiting NFκB and COX-2 and increasing the expression of P21 that plays an essential role in increasing CD14 protein expression and subsequently induce differentiation in HL-60 cells to monocytes.