title: The Evaluation and Comparison of Transcriptionally Targeted Noxa and Puma Killer Genes to Initiate Apoptosis Under Cancer-Specific Promoter CXCR1 in Hepatocarcinoma Gene Therapy.
abstract: BACKGROUND
  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.
  
  
  OBJECTIVE
  This study evaluated the potential of inducing apoptosis using Noxa and Puma in a hepatocarcinoma cell line.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran; Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, IR Iran.
authors.forename: Shahryar
authors.initials: S
authors.lastname: Khoshtinat Nikkhoi
authors.affiliation: Department of Microbiology, Faculty of Science, Azad university of Shahreh Qods, Tehran, IR Iran.
authors.forename: Hedieh
authors.initials: H
authors.lastname: Heydarzadeh
authors.affiliation: Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
authors.forename: Saeed
authors.initials: S
authors.lastname: Ranjbar
authors.affiliation: Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Salimi
authors.affiliation: Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Aghaeifard
authors.affiliation: Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, IR Iran.
authors.forename: Seyed Moayed
authors.initials: SM
authors.lastname: Alavian
authors.affiliation: Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Reshadmanesh
keywords: Cancer Gene Therapy; Killer Gene; Lentivirus; Noxa; Puma