CAJ | 학술논문

目的观察缺氧及应用小干扰RNA (siRNA)沉默缺氧诱导因子-1α(HIF-1α)基因对胰腺癌细胞株BxPC-3中RUNX3基因表达的影响.方法构建人RUNX3基因真核表达载体质粒pEGFP-RUNX3,CoC12化学模拟肿瘤缺氧环境,检测缺氧以及siRNA沉默HIF-1α对BxPC-3细胞中RUNX3基因在mRNA和蛋白水平的影响.结果缺氧能使BxPC-3细胞中RUNX3基因的表达较常氧时显著升高[表达为(9.13±1.55),P＜0.05],siRNA转染BxPC-3细胞后能显著下调HIF-1α蛋白表达(抑制率90％),并导致RUNX3基因的表达也受到明显抑制.结论缺氧促使BxPC-3细胞中HIF-1α在蛋白水平表达升高,缺氧时HIF-1α能上调RUNX3基因的表达水平.
목적 관찰결양급응용소간우RNA (siRNA)침묵결양유도인자-1α(HIF-1α)기인대이선암세포주BxPC-3중RUNX3기인표체적영향.방법 구건인RUNX3기인진핵표체재체질립pEGFP-RUNX3,CoC12화학모의종류결양배경,검측결양이급siRNA침묵HIF-1α대BxPC-3세포중RUNX3기인재mRNA화단백수평적영향.결과 결양능사BxPC-3세포중RUNX3기인적표체교상양시현저승고[표체위(9.13±1.55),P＜0.05],siRNA전염BxPC-3세포후능현저하조HIF-1α단백표체(억제솔90％),병도치RUNX3기인적표체야수도명현억제.결론 결양촉사BxPC-3세포중HIF-1α재단백수평표체승고,결양시HIF-1α능상조RUNX3기인적표체수평.
Objective To observe the expression of hypoixa inducible factor-1α (HIF-1α) and RUNX3 gene in BxPC-3 cells under hypoxia in vitro,and explore the regulation of RUNX3 gene expression by HIF-1α.Methods CoCl2 was used as a chemical hypoxia-inducible reagent to mimic tumor hypoxic microenvironment.RUNX3 mRNA and protein expression was detected.The BxPC-3 cells were transfected with RNA interference (RNAi) originated by small interference RNA (siRNA).The change of RUNX3 gene expression was observed after HIF-1α gene silencing.Results Under hypoxia for 24 h,RUNX3 in BxPC-3 cells was increased in mRNA and protein levels (P ＜0.01 ).After siRNA transfection,the HIF-1α was down-regulated efficiently in BxPC-3 cells ( inhibition ratio:90％ ),and RUNX3 gene was down-regulated as well.Conclusion Hypoxia can increase protein level of HIF-1α in BxPC-3 cells.HIF-1α up-regulates the gene expression of RUNX3 under hypoxia.