Lentiviral gene delivery is now considered as a major candidate in the future of cancer therapy. To avoid common side-effects associated with methods of cancer therapy, the survivin promoter shows great promise due to its high expression level in multiple cancers. In this research, using a 2A peptide, Noxa was coexpressed with hemagglutinin neuraminidase (HN) driven by the survivin promoter (Surp). Coding genes of Noxa and HN were connected to the upstream and downstream of a 2A peptide using SOE PCR. After cloning of Noxa-2A-HN and Surp into the lentiviral vector, the final construct was transfected to SK-BR3 tumor cells and human embryonic kidney 293T cells. Quantitative real-time PCR implied efficient transfection and integration of the lentiviral vector in target cells. Western blotting analysis was performed to verify the expression of Noxa and HN and we determined cell viability by MTT calorimetric assay after transfection. Our results showed that the expression of Noxa and HN, though not utterly specific, was relatively higher in tumor cells and led to the selective suppression of tumor cells, which eventually resulted in a cell survival reduction of 76%. In conclusion, the novel pLN2AH-Surp lentiviral vector proved the potential for further in vivo and clinical experiments.
RANJBAR, SAEED; HASHEMZADEH, MOHAMMAD SADEGH; NIKKHOI, SHAHRYAR KHOSHTINAT; FARASAT, ALIREZA; TAT, MAHDI; GHALAVAND, MAJDEDIN; and DOROSTKAR, RUHOLLAH
"Selective suppression of tumor cells by a tumor-specific bicistronic lentiviral vector,"
Turkish Journal of Biology: Vol. 40:
6, Article 18.
Available at: https://journals.tubitak.gov.tr/biology/vol40/iss6/18