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邓实, 杨博, 任正举等. 缺氧诱导因子及其抑制因子调控肾癌细胞生长的实验研究[J]. koko体育app 学报（医学版）, 2018, 49(1): 29-33.
引用本文: 邓实, 杨博, 任正举等. 脑缺氧帮助因素举例说明抑制作用因素国家宏观调控肾癌上皮细胞出现的研发研发[J]. 广东大学时学报（中医药学版）, 2018, 49(1): 29-33.
DENG Shi, YANG Bo, REN Zheng-ju. et al. Growth Regulation of Factor Inhibiting Hypoxia-Inducible Factor in Renal Carcinoma Cells[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(1): 29-33.
Citation: DENG Shi, YANG Bo, REN Zheng-ju. et al. Growth Regulation of Factor Inhibiting Hypoxia-Inducible Factor in Renal Carcinoma Cells[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(1): 29-33.

缺氧诱导因子及其抑制因子调控肾癌细胞生长的实验研究

Growth Regulation of Factor Inhibiting Hypoxia-Inducible Factor in Renal Carcinoma Cells

  • 摘要: 目的 检测缺氧诱导因子抑制因子（FIH-1）在肾癌细胞中的基因和蛋白表达情况，以进一步了解FIH-1对肾癌细胞生长调控的机制。方法 选取人肾癌细胞株786-0及OS-RC-2，运用real-time PCR（RT-PCR）和Western blot方法检测细胞株中FIH-1、缺氧诱导因子1α（HIF- 1α）、 缺氧诱导因子2α（HIF- 2α）、血管内皮生长因子（VEGF）的表达水平。并人工构建FIH-1过表达质粒，将其转入上述的两种细胞株中，运用RT-PCR和Western blot方法观察转染后FIH-1、HIF及VEGF的表达情况；同时运用MTT方法分析转染后两种肿瘤细胞的增殖情况。结果 RT-PCR和Western blot检测结果都显示，786-0和OS-RC-2细胞中都能检测到FIH-1和VEGF的表达；OS-RC-2细胞中同时存在HIF-1α和HIF-2α表达，而786-0细胞中只有HIF-2α的表达；786-0细胞中FIH-1表达比OS-RC-2细胞高，VEGF的表达比OS-RC-2低; 转染FIH-1过表达质粒后，OS-RC-2细胞内的FIH-1表达量明显升高，下游蛋白HIF-1α的表达量降低，VEGF的表达量也降低，而HIF-2α的表达量前后不变；在786-0细胞内的FIH-1表达量明显升高时，HIF-2α的表达量不变，VEGF的表达量也不变。在OS-RC-2细胞中，转染FIH-1过表达质粒后细胞的增殖变缓，而786-0细胞的增殖则没有明显的变化。结论 在肾癌细胞中存在FIH-1/HIF-1α/VEGF信号通路，FIH-1反向调节HIF-1α的表达，HIF-1α正向调节VEGF的表达。在HIF-1α及HIF-2α都存在时，FIH-1可通过对HIF-1α调控影响HIF-1α下游基因表达，抑制肾癌细胞的增殖。  
    Abstract: Objective To detect the expression and growth regulation effects of the factor inhibiting hypoxia-inducible factor (FIH-1) in renal carcinoma cells. Methods The expressions of FIH-1, hypoxia inducible factor (HIF) and vascular endothelial growth factor (VEGF) were examined in OS-RC-2 and 786-0 cell lines via real-time PCR (RT-PCR) and Western blot. Overexpression plasmid of FIH-1 were designed and constructed as EX-V1104-M45-FIH-1. After the transfection of the plasmid into OS-RC-2 and 786-0 cells, the expression levels of FIH-1, HIF and VEGF were measured by RT-PCR and Western blot, while cell proliferation was tested by MTT. Results The expressions of FIH-1, HIF-1α, HIF-2α and VEGF were detected in OS-RC-2 cells, and FIH-1, HIF-2α and VEGF were detected in 786-0 cells. After the transfection of EX-V1104-M45-FIH-1,the level of FIH-1 was increased in OS-RC-2, while the levels of HIF-2α, HIF-1α and VEGF were decreased. MTT showed that FIH-1 plasmid transfection decreased the cell proliferation of OS-RC-2, but did not show inhibition on 786-0 cells proliferation. Conclusion FIH-1/HIF-1α/VEGF signaling pathway exists in renal carcinoma cell lines.  
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