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| DOI： |
| 中文关键词: 缺氧诱导因子 糖酵解 结肠癌 |
| 英文关键词:hypoxia%20inducible%20factor-1α %20glycolysis%20 colon%20carcinoma |
| 基金项目:鄂科技发计[2014]11号 2014CC1040 |
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| 摘要点击次数: 149 |
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| 中文摘要: |
| 目的：%20探讨RNA干扰沉默缺氧诱导因子(HIF-1α)对缺氧环境下培养的结肠癌细胞糖酵解的影响。方法：%20选择人结肠癌SW480细胞稳定转染HIF-1αSiRNA及空质粒细胞,予以缺氧环境培养,分为3组：空白对照组、空质粒细胞组、HIF-1αSiRNA组,培养时间为24小时。采用RT-PCR和Western-blot法检测各组结肠癌细胞HIF-1αmRNA和蛋白表达水平；RT-PCR和Western-blot法检测结肠癌细胞糖酵解相关基因(GLUT-1,LDH-A)的表达。比色法测定各组结肠癌细胞上清液中乳酸的浓度。结果：%20缺氧环境下,SiRNA抑制结肠癌细胞SW480中HIF-1α表达后,糖酵解相关酶GLUT-1、LDH-A下降(P＜0.05),结肠癌细胞培养上清液中乳酸浓度降低(P＜0.05)。结论：%20缺氧条件下,利用RNA干扰沉默HIF-1α表达后,可导致糖酵解相关基因GLUT-1、LDH-A的表达下降和乳酸含量的下降(P＜0.05),提示HIF-1α参与了结肠癌细胞的糖酵解过程。 |
| 英文摘要: |
| Objective:%20To%20study%20the%20effect%20of%20RNA%20interference-based%20silencing%20hypoxia-inducible%20factor%201α(HIF-1α)%20silencing%20on%20the%20expression%20%20of%20%20glycosis%20in%20colon%20carcinoma%20cells.%20%20Methods:%20%20Colon%20carcinoma%20cells(SW-480)%20were%20cultured%20in%20a%20hypoxia%20environment.SW-480%20cells%20were%20divided%20into%20three%20groups%20,namely,untransfected%20%20SW480%20cells,SW480%20cells%20stably%20transfected%20with%20HIF-1αSiRNA%20and%20empty%20expression%20vector.After%20the%20cells%20were%20cultured%20in%20a%20hypoxia%20environment%20for%2024h,the%20expression%20of%20%20HIF-1α%20protein%20was%20detected%20by%20western-blott,and%20the%20expression%20of%20glycolysis%20associated%20genes(GLUT-1,LDH-A)%20%20was%20detected%20by%20RT-PCR%20and%20western-blotting.%20colorimetric%20method%20was%20employed%20to%20determine%20the%20the%20concentration%20of%20%20lactic%20.Results:%20After%20SW480%20cells%20were%20cultured%20in%20a%20hypoxia%20environment,the%20expression%20of%20%20HIF-1α%20was%20upregulated,while%20the%20expression%20of%20HIF-1α%20was%20hardly%20detected%20in%20HIF-1α%20silence%20cells.The%20concentration%20of%20%20lactic%20was%20obviously%20decreased%20in%20HIF-1α%20silence%20cells. |
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