Adenovirus-mediated co-expression of the TRAIL and HN genes inhibits growth and induces apoptosis in Marek's disease tumor cell line MSB-1

Cancer Cell International
Dongxiao DongTianming Jin

Abstract

The objective of this study was to determine the in vitro tumor-inhibitory effect of a recombinant adenovirus expressing a fusion protein of tumor necrosis factor (TNF) related apoptosis inducing ligand (TRAIL) and hemagglutinin-neuraminidase (HN) genes on the MSB-1 Marek's disease tumor cell line. TRAIL and HN genes were amplified from lymphocytes in the peripheral blood of chickens and the LaSota strain of Newcastle disease virus (NDV), respectively, using RT-PCR. The two genes were connected with a 2A connecting peptide by site-directed mutagenesis and gene splicing by overlap extension (SOE). The target gene TRAIL-2A-HN was cloned into the shuttle vector pShuttle-CMV. Homologous recombination was carried out with the vector pAdeasy-1 in the bacterium BJ5183 to construct the recombinant adenovirus plasmid pAd-TRAIL-2A-HN. After linearization, the plasmid was transfected into AD293 cells and packaged. Real-time quantitative PCR (RT-PCR) and fluorescence microscopy confirmed the introduction of the recombinant adenovirus into AD293 cells. The TCID50 method (50% tissue culture infectious dose) was employed to determine viral titers for the exprimental and control viruses, which met criteria for use. The Marek's disease tumor ce...Continue Reading

References

Apr 20, 1999·The Journal of Experimental Medicine·T S GriffithN A Fanger
Jul 1, 1965·Cancer·W A CASSEL, R E GARRETT
Nov 20, 2004·Cancer Gene Therapy·Lidong Zhang, Bingliang Fang
Jan 16, 2007·Molecular Cancer Therapeutics·Hiroaki ItamochiNaoto T Ueno
Jan 15, 2009·Results and Problems in Cell Differentiation·Stefanie M CordierHenning Walczak
Mar 2, 2010·Cancer Biotherapy & Radiopharmaceuticals·Jisheng LiuJicheng Yang
Oct 6, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Tokunori IkedaSatoru Senju
Aug 1, 2012·Avian Pathology : Journal of the W.V.P.A·Peter M Biggs, Venugopal Nair

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
transfect
transfection
density gradient centrifugation
electrophoresis
ELISA
flow cytometry

Software Mentioned

SOE

Related Concepts

Related Feeds

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

Related Papers

Sheng wu gong cheng xue bao = Chinese journal of biotechnology
Shuangjuan YangGongshe Yang
Journal of Huazhong University of Science and Technology. Medical Sciences = Hua Zhong Ke Ji Da Xue Xue Bao. Yi Xue Ying De Wen Ban = Huazhong Keji Daxue Xuebao. Yixue Yingdewen Ban
Qingyong ChenHai Zheng
Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi
Dong XiaLanying Zhao
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Kailing JiangBeizhong Liu
© 2022 Meta ULC. All rights reserved