Lentivirus-mediated RNA interference targeting E2F-1 inhibits human gastric cancer MGC-803 cell growth in vivo.
10.3858/emm.2011.43.11.072
- Author:
Xiao Tong WANG
1
;
Yu Bo XIE
;
Qiang XIAO
Author Information
1. Department of Surgery, The First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China. xiaoqiang20050@yahoo.com.cn
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
E2F1 transcription factor;
gene therapy;
lentivirus;
RNA interference;
stomach neoplasms
- MeSH:
Animals;
Apoptosis;
Blotting, Western;
Caspase 3/metabolism;
Caspase 9/metabolism;
Cells, Cultured;
E2F1 Transcription Factor/antagonists & inhibitors/genetics/*metabolism;
Genetic Vectors/administration & dosage;
Humans;
Lentivirus/*genetics;
Male;
Mice;
Mice, Inbred BALB C;
Mice, Nude;
RNA Interference;
RNA, Small Interfering/*genetics;
Stomach Neoplasms/*genetics/pathology/*prevention & control
- From:Experimental & Molecular Medicine
2011;43(11):638-645
- CountryRepublic of Korea
- Language:English
-
Abstract:
The E2F-1 transcription factor is post-translationally modified and stabilized in response to various forms of DNA damage to regulate the expression of cell-cycle and pro-apoptotic genes. The sustained overexpression of E2F-1 is a characteristic feature of gastric cancer. In this study, we investigated the role of short hairpin RNA (shRNA) targeting E2F-1 gene on human gastric cancer MGC-803 cell growth in vivo, and preliminarily revealed the mechanism. Thus, we constructed recombinant pGCSIL-GFP-shRNA-E2F-1 lentiviral vector to knock down E2F-1 expression in human gastric cancer MGC-803 cells in vivo, and studied the effect of E2F-1 shRNA on growth of MGC-803 tumor and evaluated its treatment efficacy. Our data demonstrated that in a mouse model of established gastric cancer, intratumor injection of lentiviral shRNA targeting E2F-1 definitely decreased the endogenous E2F-1 mRNA and protein expression in MGC-803 tumor, and inhibited tumor growth and promoted tumor cells apoptosis. Moreover, we found that E2F-1 shRNA increased the expression of phosphatase and tensin homolog (PTEN), activated caspase-3 and caspase-9, and suppressed nuclear factor (NF)-kappaB expression in tumor tissue as determined by reverse transcription (RT)-PCR and western blotting. In summary, shRNA targeting of E2F-1 can effectively inhibits human gastric cancer MGC-803 cell growth in vivo and may be a potential therapeutic strategy for gastric cancer.
