The Evaluation and Comparison of Transcriptionally Targeted Noxa and Puma Killer Genes to Initiate Apoptosis Under Cancer-Specific Promoter CXCR1 in Hepatocarcinoma Gene Therapy
| Author | Shahryar Khoshtinat Nikkhoi | en |
| Author | Hedieh Heydarzadeh | en |
| Author | Saeed Ranjbar | en |
| Author | Fatemeh Salimi | en |
| Author | Masoud Aghaeifard | en |
| Author | Seyed Moayed Alavian | en |
| Author | Azadeh Reshadmanesh | en |
| Orcid | Seyed Moayed Alavian [0000-0002-4443-6602] | en |
| Issued Date | 2016-10-01 | en |
| 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. Objectives: 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. | en |
| DOI | https://doi.org/10.5812/hepatmon.38828 | en |
| Keyword | Lentivirus | en |
| Keyword | Killer Gene | en |
| Keyword | Puma | en |
| Keyword | Noxa | en |
| Keyword | Cancer Gene Therapy | en |
| Publisher | Brieflands | en |
| 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 | en |
| Type | Research Article | en |