In-vitroCharacterization and Evaluation of Apoptotic Potential of Bicistronic Plasmid Encoding HN Gene of Newcastle Disease Virus and Human TNF-α
| Autor: | Lakshya Veer Singh, Rajiv Kumar, Shishir Kumar Gupta, Aditya Prasad Sahoo, Shikha Saxena, R. S. Rajmani, Prafull Kumar Singh, G.V.P.P.S. Ravi Kumar, Ashok K. Tiwari, Uttara Chaturvedi |
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| Rok vydání: | 2014 |
| Předmět: |
HN Protein
Expression vector Tumor Necrosis Factor-alpha Genetic enhancement Genetic Vectors Newcastle disease virus Apoptosis Bioengineering Genetic Therapy Transfection Biology biology.organism_classification Molecular biology Recombinant Proteins Virus HeLa Plasmid Gene expression Multiple cloning site Humans Animal Science and Zoology HeLa Cells Plasmids Biotechnology |
| Zdroj: | Animal Biotechnology. 26:112-119 |
| ISSN: | 1532-2378 1049-5398 |
| Popis: | The viral gene oncotherapy in combination with cytokines emerges as an exciting strategy for cancer therapy due to its minimal side effects and tumor specificity. HN is the surface protein of NDV which is involved in virus infectivity and is known to kill many cancerous cell types. TNF-α, a multifactorial cytokine has direct anti-tumor activity by activating the extrinsic pathways of apoptosis. In the present study, HN gene of NDV and TNF-α of human were cloned at multiple cloning sites (MCS) 1 and 2 of bicistronic expression vector pVIVO2. Expression pattern of recombinant clone was checked on transcriptional and translational level by RT-PCR, Immunofluorescence assay and flow cytometry. On flow cytometric analysis HN gene expression was found to be 28.30 ± 1.21; 5.22 ± 0.60%, and TNF-α gene expression was found to be 15.44 ± 0.42; 6.51 ± 0.757%, in HeLa cells transfected with pVIVO.nd.hn.hu.tnf and pVIVO2 empty vector control, respectively. These assays confirm that HN and TNF-α act synergistically in the induction of apoptosis in HeLa cells. |
| Databáze: | OpenAIRE |
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