Effects of Apis dorsata Honey on the mRNA Expression of Selected CYP450, Pro-apoptotic, and Anti-apoptotic Genes during Induced Cytotoxicity in Cyclophosphamide-treated Human Lung Carcinoma (A549) Cells.

Abstract

Introduction: One of the novel strategies in cancer treatment is the combination of conventional chemotherapeutic drugs and natural products. In a previous study, co-treatment of the anti-cancer drug cyclophosphamide (CP) with honey from giant honey bee (Apis dorsata) resulted to a dose-dependent increase in its cytotoxic effect in human lung carcinoma (A549) cells. However, the molecular mechanism of this combinatorial effect remains unknown.

Objectives: In this study, the effect of A. dorsata honey on the expression of selected CYP450 genes at the mRNA level, as well as the proapoptotic gene CASP8 and antiapoptotic gene BCL2 was investigated in CP-treated A549 cells.

Methods: MTT Assay was performed to determine the cell viability of A549 cells after treatment with CP with or without A. dorsata honey, as well as the EC₅₀ of CP with honey thereafter. RT-qPCR was then performed to study the effect of A. dorsata honey on the expression of selected CYP450 genes as well as CASP8 and BCL2 genes in CP-treated A549 cells. LC-MS was carried out to screen for putative compounds in A. dorsata honey which may possibly have anti-cancer activity.

Results: Honey in the lowest concentration (0.6% v/v) most effectively enhanced the cytotoxic effect of CP. CYP2J2 and CYP1B1 indicated a 2.38-fold and 1.49-fold upregulation, respectively as compared to untreated cells. This cytotoxic effect is further enhanced by upregulation of CASP8 that is paralleled by a downregulation of BCL2. Phytosphingosine and sphinganine are honey constituents which may be linked to the increased cytotoxicity of CP observed in A549 cells.

Conclusion: This study provides further knowledge on the molecular basis by which A. dorsata honey potentiates the cytotoxic effect of cyclophosphamide in A549 cells.