مهار ژن هومانین جهت بررسی اثرات آن در اثر بخشی داروی Etoposide در رده سلولی سرطان معده

Background and purpose: Humanin is a 24-aa peptide that is considered as one of the probable players of autophagy. Its inevitable role in the inhibition of apoptosis in cells is revealed, but its probable importance in autophagy is under evaluation. This study evaluated the role of humanin protein in induction of autophagy and efficacy of chemotherapy drug etoposide. Materials and methods: HN3 gene was synthesized and cloned in pcDNA3.1 vector and electroporated in AGS cell line. Expression analysis in AGS cell line was done by SDS-PAGE and Real-time PCR using specific primers. HN3 gene inhibition was done using specific siRNA molecules. siRNAs transfection to the recombinant AGS cell line was done using Lipofectamine. The effects of the humanin inhibition on efficacy of chemotherapy agents was studied using Real-time PCR and MTT assay. Results: Our data showed that dose of 100 nm of siRNA caused about 40% increase in the mortality of cancer cells. Conclusion: To the best of our knowledge the current research was the first on the role of humanin gene in induction of autophagy in stressful cancer cells that evaluated its exact mechanism in autophagy induction. In addition, HN3 inhibition could increase the efficacy of chemotherapy agents in cancer cells.