Author/Authors :
Khoshtinat Nikkhoi، Shahryar نويسنده Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran , , Heydarzadeh، Hedieh نويسنده Department of Microbiology, Faculty of Science, Azad university of Shahreh Qods, Tehran, IR Iran , , Ranjbar، Saeed نويسنده Institute of Cardiovascular Research, Modarres Hospital, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran , , Salimi، Fatemeh نويسنده Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran , , Aghaeifard، Masoud نويسنده Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran , , Alavian، Seyed-Moayed نويسنده , , Reshadmanesh، Azadeh نويسنده Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran ,
Abstract :
Cancerous cells proliferate as fast as possible without a proper surveillance system. This rapid cell division leads to enormous mutation rates, which help a tumor establish. This study evaluated the potential of inducing apoptosis using Noxa and Puma in a hepatocarcinoma cell line. The current study generated two recombinant lentiviruses, pLEX-GCN and pLEX-GCP, bearing Noxa and Puma, respectively. Transduction of both genes to hepatocarcinoma (HepG2) was verified using fluorescent microscopic analysis, western blotting, and quantitative real-time polymerase chain reaction (PCR). To evaluate the potential of Noxa and Puma to initiate apoptosis, a caspase-9 real-time, MTT assay, and a 4’, 6-diamidino-2-phenylindole (DAPI) reagent were performed to stain apoptotic cells. The data verified successful transduction to HepG2 and HEK293T. Higher relative expression of Noxa and Puma rather than the untransduced cell line showed these genes are expressed more in HepG2 in comparison to HEK293T. The results of the real-time PCR, MTT assay, and DAPI reagent illustrated that higher cells initiated apoptosis following Puma transduction rather than Noxa. In this approach, the suicide gene was transferred to transformed cells and ignited apoptosis to exterminate them. Puma is a more potent killer gene and has higher capabilities to start intrinsic apoptosis pathway.
