The Effect of Honey Bee Venom on CD۱۴ Protein Expression as Monocyte Marker in D-Alpha-Tocopheryl Succinate (Vitamin E)-Treated HL-۶۰ Cells

Background: The membrane form of a cluster of differentiation ۱۴ (CD۱۴) 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 HL-۶۰ cell surface. It is believed that the differentiation of HL-۶۰ cells stops in the promyelocytic stage. The differentiation of HL-۶۰ cells with compounds such as vitamin A, D, E results in membrane CD۱۴ expression. Objectives: The objective of this study was an evaluation of CD۱۴ expression by Honey Bee Venom (HBV) in HL۶۰ cells treated by D-alpha-tocopheryl succinate (D-α-TS). Methods: HL-۶۰ cells were cultured in RPMI Media ۱۶۴۰ 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 ۳ software. Methods: HL-۶۰ 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 ۳ software. Results: MTT assay demonstrated that HBV and D-alpha-tocopheryl induce death in HL-۶۰ 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 ۶ µg/mL D-α-TS and ۲.۵ µg/mL HBV for ۵ days significantly increase the expression of CD۱۴ in HL-۶۰ 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 NF-κB and COX-۲ and increasing the expression of P۲۱ that plays an essential role in increasing CD۱۴ protein expression and subsequently induce differentiation in HL-۶۰ cells to monocytes.​​​​​​

Keywords:

Differentiation , Hematopoietic stem cell , Promyelocytic leukemia , p۲۱ protein