Please use this identifier to cite or link to this item: http://nopr.niscpr.res.in/handle/123456789/31435
Title: Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1
Authors: Saxena, Shikha
Desai, GS
Kumar, G Ravi
Sahoo, AP
Santra, Lakshman
Singh, Lakshya Veer
Kumar, Sudesh
Tiwari, AK
Keywords: Apoptosis;Cancer;Canine Parvovirus-2 (CPV-2);Confocal microscopy;DNA fragmentation;Flowcytometry;Oncotherapy
Issue Date: May-2015
Publisher: NISCAIR-CSIR, India
Abstract: Viral 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.
Page(s): 249-255
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.53(05) [May 2015]

Files in This Item:
File Description SizeFormat 
IJEB 53(5) 249-255.pdf400.92 kBAdobe PDFView/Open


Items in NOPR are protected by copyright, with all rights reserved, unless otherwise indicated.