Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

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dc.contributor.authorSaxena, Shikha
dc.contributor.authorDesai, G S
dc.contributor.authorKumar, G Ravi
dc.contributor.authorSahoo, A P
dc.contributor.authorSantra, Lakshman
dc.contributor.authorSingh, Lakshya Veer
dc.contributor.authorKumar, Sudesh
dc.contributor.authorTiwari, A K
dc.date.accessioned2015-06-26T07:08:29Z
dc.date.available2015-06-26T07:08:29Z
dc.date.issued2015-05
dc.description.abstractViral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.en_US
dc.identifier.citationSaxena Shikha, Desai G S, Kumar G Ravi, Sahoo A P, Santra Lakshman, Singh Lakshya Veer, Kumar Sudesh, Tiwari A K. Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1. Indian Journal of Experimental Biology. 2015 May; 53(5): 249-255.en_US
dc.identifier.issn0975-1009
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/158441
dc.language.isoenen_US
dc.publisherNISCAIR-CSIR, Indiaen_US
dc.source.urihttps://nopr.niscair.res.in/handle/123456789/31435en_US
dc.subjectApoptosisen_US
dc.subjectCanceren_US
dc.subjectCanine Parvovirus-2 (CPV-2)en_US
dc.subjectConfocal microscopyen_US
dc.subjectDNA fragmentationen_US
dc.subjectFlowcytometryen_US
dc.subjectOncotherapyen_US
dc.subject.meshApoptosis
dc.subject.meshCell Cycle --analysis
dc.subject.meshCell Cycle --genetics
dc.subject.meshDNA Fragmentation
dc.subject.meshFlow Cytometry --methods
dc.subject.meshGenes, Viral --genetics
dc.subject.meshMicroscopy, Confocal --methods
dc.subject.meshNeoplasms --therapy
dc.subject.meshParovirus, Canine --genetics
dc.titleCharacterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.en_US
dc.typeArticleen_US
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