Evaluation of E1B-mutant Replicating Adenoviruses for Cancer Gene Therapy.
- Author:
Jae Sung KIM
1
;
Joo Hang KIM
;
Heui Ran LEE
;
Kyeong Cheon JUNG
;
Chae Ok YUN
Author Information
1. Institute of Cancer Research, Yonsei Cancer Center, Yonsei University College of Medicine, Seoul, Korea. chaeok@yumc.yonsei.ac.kr
- Publication Type:Original Article
- Keywords:
Cancer gene therapy;
Replication-competent adenovirus;
Apoptosis
- MeSH:
Adenoviridae*;
Apoptosis;
Cellular Structures;
DNA;
DNA Fragmentation;
Genes, Neoplasm*;
In Situ Nick-End Labeling;
Nuclear Envelope;
Organelles
- From:Cancer Research and Treatment
2001;33(6):500-511
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Gene-attenuated replication-competent adenoviruses are emerging as a promising new modality for the treatment of cancer. In an effort to continually improve upon cancer gene therapy, we have modified gene- attenuated replication-competent adenoviruses so as to cause them to replicate efficiently and lyse the infected cancer cells more effectively. MATERIALS AND METHODS: We modified the E1 region of the adenovirus (Ad) systematically, generating Ad-deltaE1B19, Ad-deltaE1B55, Ad-deltaE1B19/55, and Ad-WT. The cytopathic effects (CPE) and viral replication of these four gene modified adenoviruses were compared, and the morphology and DNA fragmentation of the infected cells was evaluated. RESULTS: Among the constructed adenoviruses, E1B 19kD-inactivated adenovirus (Ad-deltaE1B19) was the most potent, inducing the largest-sized plaques and markedCPE. Moreover, cells infected with Ad-deltaE1B19 showed complete cell lysis with disintegrated cellular structure whereas cells infected with Ad-WT maintained intact cellular and nuclear membrane with properly structured organelles. TUNEL assay was also used to monitor DNA integrity, and a more profound induction of apoptosis was observed in the Ad-deltaE1B19 infected cells in comparison to wild type adenovirus infected cells. CONCLUSION: We demonstrate that the inactivation of the E1B19kD gene in a replicating adenovirus leads to increased CPE, rapid viral release, improved cell-to-cell viral spread and increased induction of apoptosis.
