Construction of RNAi targeting TRAF1 gene and effect of TRAF1 on gastric cancer cells.
- Author:
Fen WANG
1
;
Yan YANG
;
Qian FENG
;
Guangkui BU
;
Lihua HUANG
;
Hongwei LU
;
Qin GUO
;
Canxia XU
;
Shourong SHEN
Author Information
1. Department of Gastroenterology, Third Xiangya Hospital, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
genetics;
Base Sequence;
Cell Line, Tumor;
Humans;
Molecular Sequence Data;
RNA Interference;
RNA, Small Interfering;
genetics;
Stomach Neoplasms;
genetics;
metabolism;
pathology;
TNF Receptor-Associated Factor 1;
genetics;
metabolism;
Transfection;
Up-Regulation;
genetics
- From:
Journal of Central South University(Medical Sciences)
2012;37(9):876-882
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct the RNAi targeting tumor necrosis factor receptor associated factor (TRAF1) gene, and to explore the effect of interference targeting TRAF1 on the biological behavior of gastric cancer cells.
METHODS:We detected the expression of TRAF1 in BGC823, SGC7901, and MGC803 gastric cancer cell lines through the real-time PCR and Western blot; then we constructed three pLVXshRNA- TRAF1-shRNAs expression vector targeting TRAF1. When TRAF1 was interfered successfully, we selected the strongest interference efficiency ShRNA by real-time PCR and Western blot. Based on interference targeting TRAF1 on gastric cancer, we tested the cell proliferation activity and apoptosis through MTT assay and flow cytometry, and the cell migration by transwell migration assay.
RESULTS:The expression of TRAF1 was increased in BGC823, SGC7901, and MGC803 gastric cancer cell lines compared with gastric epithelial cells (P<0.05), and the highest expression was in BGC823 gastric cell line. In the three TRAF1 shRNAs, the strongest interference efficiency shRNA was pLVX-shRNA-TRAF1-shRNA2. When the gene TRAF1 of BGC823 was interfered, the cell growing power was weakened and the apoptosis rate increased, and the cell migration had no difference.
CONCLUSION:The expression of TRAF1 is up-regulated in gastric cancer cell lines BGC823, SGC7901, and MGC803, and the most obvious one is BGC823. The interference targeting TRAF1 can successfully inhibit the expression of TRAF1 in gastric cancer cell line BGC823. TRAF1 can inhibit the apoptosis of BGC823 cells.
