中国临床医学2012，Vol.19Issue(6)：598-601,4.
RNA干扰HIF-2α基因对人肺腺癌A549细胞株侵袭转移的影响
Effect of RNA Interference Targeting HIF-2α Gene on Invasion and Proliferation of Lung Adenocarcinoma A549 Cell Line
摘要
Abstract
Objective:To investigate the effect of small interfering RNA (siRNA) targeting hypoxia-inducible factor 2α(HIF-2α) gene on the invasion and metastasis of A549 cell line. Methods:The siRNA eukaryotic expression vectors targeting HIF-2a gene were designed and transfected into A549 cell line. Quantitative real-time PCR (qRT-PCR) and Western blot were used to detect mRNA and protein expression of HIF-2α gene. The migration of A549 cells was assayed using transwell ceil culture chambers, and vascular endothelial growth factor(VEGF) protein expression was determined by enzyme-linked immunosorbent assay (ELISA). Results: After transfection of HIF-2crsiRNA into A549. mRNA and protein expression of HIF-2α were down-regulated. The results of qRT-PCR showed that No. 4 siRNA vector was the most effective one in suppressing HIF-2a mRNA expression. Transfection of A549 with this siRNA vector resulted in sequence-specific silencing with decreases of (60. 63 ± 5. 10)% and (80. 00 ± 3. 55)% in HIF-2α mRNA transcription at 24h or 48h posMransfection. The Western blot test also demonstrated the best effectiveness of this expression vector with the inhibition rates of (31. 69 ± 11. 56) % at 24h post-trans-fection and (82. 9 ± 4. 09) % at 48h post-transfection. It was shown in migration analysis that the number of cells which had migrated through the filter was much amaller in groups treated with HIF-2α-siRNA than in control, which indicated a reduction in the invasive capacity of this group. ELISA analysis showed the protein expression of VEGF was significantly down-regulated when A549 cells were treated with siRNA targeting HIF-2α transfection. Conclusions: Eukaryotic expression vectors of siRNA which inhibit the expression of HIF-2α gene can effectively suppress the invasion and metastasis of A549 cells.关键词
肺腺癌/缺氧诱导因子-2α/小干扰RNA/侵袭转移Key words
Lung adenocarcinoma/ Hypoxia-inducible factor 2a/ Small interfering RNA/ Invasion and metastasis分类
医药卫生引用本文复制引用
袁凯,王勇,童继春,袁卫东,陈栋..RNA干扰HIF-2α基因对人肺腺癌A549细胞株侵袭转移的影响[J].中国临床医学,2012,19(6):598-601,4.基金项目
江苏省常州市卫生局青年基金(编号:QN201003) （编号:QN201003）
