Exp Mol Med.  2011 Nov;43(11):638-645. 10.3858/emm.2011.43.11.072.

Lentivirus-mediated RNA interference targeting E2F-1 inhibits human gastric cancer MGC-803 cell growth in vivo

• Wang XT
• Xie YB
• Xiao Q

• KMID: 1025167
• DOI: http://doi.org/10.3858/emm.2011.43.11.072

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.

Keyword

E2F1 transcription factor; gene therapy; lentivirus; RNA interference; stomach neoplasms

MeSH Terms

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