85 Summer & Autumn 2021, Volume 18, Number 2 Research Paper: The Efects of Melittin Coding Gene of Bee Venom on Bcl-2 and Bax Genes Expression in ACHN Cells Erfaneh Heidari Esfahani 1 , Abbas Doosi 2 * 1. Department of Biology, School of Science, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran. 2. Biotechnology Research Center, Shahrekord Branch Islamic Azad University, Shahrekord, Iran. * Corresponding Author: Abbas Doosi Address: Biotechnology Research Center, Shahrekord Branch Islamic Azad University, Shahrekord, Iran. Tel: +98 (913) 3838830 E-mail: abbasdoosi@yahoo.com A B S T R A C T Keywords: ACHN cell line, Bcl-2, Bax, Melittin, Apoptosis Introduction: Venom therapy is a traditional procedure for cancer treatment. Various types of natural compositions in bee venom indicate an area of research for therapeutic and anti- cancer purposes. Melittin is a cationic and amphipathic peptide that interferes with lipid-protein molecular interactions in cell membranes leading to cell lysis. Melittin also prevents metasasis in hepatocarcinoma through inhibition of cell migration. This sudy aimed to invesigate the expression of Bcl-2 and Bax genes in ACHN cells transfected with a recombinant vector carrying bee melittin gene by real-time RT-PCR technique. Methods: In the present sudy, the recombinant plasmid of pcDNA3.1(+)-melittin was designed. Then, the synthesized gene was cloned into the pcDNA3.1(+) vector. Following plasmid amplifcation, PCR assay and enzymatic digesion method were used to approve the accuracy of plasmid amplifcation. ACHN cells were cultured, and sham and control groups were also prepared. RNA was extracted from the treatment and control groups using RNX-Plus solution. cDNA was generated using a specifc laboratory kit. A real-time RT-PCR assay was performed to evaluate the expression of Bcl-2 and Bax genes in ACHN cells transfected with recombinant vector. Finally, the obtained data were analyzed using SPSS. Results: The expression of the Bax gene signifcantly (P=0.03) increased in pcDNA3.1(+)- melittin compared to the pcDNA3.1(+) group. Bcl-2 gene expression showed a signifcant decreasing trend in the experimental group of pcDNA3.1(+)-melittin (P=0.001) compared to the pcDNA3.1(+). Conclusion: According to the obtained results, the expression of Bax and Bcl-2 genes in the ACHN cell line can be afected by the melittin gene of bee venom. Thus, bee venom as a natural subsance with its anti-cancer properties can be considered for further experimental invesigations. Article info: Received: 10 Feb 2021 Accepted: 23 May 2021 Available Online: 01 Jul 2021 Citation Heidari Esfahani E, Doosi A. The Efects of Melittin Coding Gene of Bee Venom on Bcl-2 and Bax Genes Expres- sion in ACHN Cells. Anatomical Sciences. 2021; 18(2):85-92. Use your device to scan and read the artcle online