Effects of lentiviral vector-mediated RNA interference of hypoxia inducible factor-1α on the expression of related genes in human pancreatic cancer Patu8988 cells
10.3760/cma.j.issn.2095-2848.2013.05.015
- VernacularTitle:慢病毒载体介导HIF-1α RNAi对人胰腺癌细胞Patu8988相关基因表达的影响
- Author:
Guanglei FAN
;
Minbin DENG
;
Yiwei WU
;
Qixian YANG
;
Bin ZHANG
;
Longbao XU
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
Tumor cells,cultured;
Gene expression;
Hypoxia-inducible factor 1,alpha subunit;
RNA interference
- From:
Chinese Journal of Nuclear Medicine and Molecular Imaging
2013;33(5):377-380
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effect of the lentiviral vector (LV)-mediated RNA interference (RNAi) targeting HIF-1α on the expression of HIF-1α and Glut-1 in human pancreatic cancer Patu8988 cells.Methods The RNAi targeting HIF-1α was combined to LV,and transfected into Patu8988 cells.The Patu8988 cells transfected with the empty vector and exposed to 0.5％ O2 for 4 h served as hypoxia negative control,the Patu8988 cells not transfected with vector and exposed to 0.5％ O2 for 4 h as hypoxia blank control,and the Patu8988 cells transfected with LV-RNAi-HIF-1α and exposed to 0.5％O2 for 4 h as experimental group.The expression of HIF-1α was measured by RT-PCR and Western blot respectively.The expression of Glut-1 was measured by RT-PCR.Each group was compared according to oneway analysis of variance and two-sample t test.Results After transfection with LV-RNAi-HIF-1α,HIF-1α mRNA expression decreased by 65.1％ (0.209/0.321) and 80.6％ (0.791/0.982) (t=10.52,15.24,both P＜0.05) under normoxia and hypoxia conditions,meanwhile with the empty vector,HIF-1α mRNA expression decreased by 0.6％ (0.002/0.321) and 7.2％ (0.071/0.982) (t =5.26,7.38,both P＜0.05).Under hypoxia conditions,the protein of HIF-1α in experimental group cells (0.159±0.010) was down-regulated obviously compared to the negative control group (0.745± 0.012) and the blank control group (0.711 ± 0.023)(F=35.52,t =6.72,10.56,all P＜0.05).The expression of Glut-1 mRNA in experimental group cells (0.040±0.003) decreased obviously compared to the negative control group (0.054±0.003) and blank control group (0.062±0.004) (F=35.28,t=5.94,8.55,all P＜0.01).Conclusion Gene silencing of HIF-1α using LV-mediated RNAi can inhibit the expression of HIF-1α and decrease the expression of Glut-1mRNA in Patu8988 cells.
