缺氧对肾癌细胞STC1、HIF-1α及线粒体膜电势稳定影响的研究

Study on the expression of STC1, HIF-1α and the effect of mitochondrial membrane potential stability in renal carcinoma cell line under hypoxic condition

目的检测缺氧条件下肾癌细胞斯钙素蛋白1(STC1)及线粒体膜电势稳定指标的变化,结合细胞增殖和Ca2+水平,初步探讨肾癌细胞可能的抗乏氧机制。方法分别使用50μmol/L、100μmol/L、150μmol/L的CoCl2干预细胞培养基构建化学性缺氧模型;MTT法检测细胞生长情况,荧光分光光度法检测细胞内Ca2+水平和线粒体膜电位(ΔΨm),紫外分光光度计测MPTP,RT-PCR检测STC1、HIF-1α的基因表达情况。结果与对照组相比,缺氧能显著抑制细胞增殖,促进细胞内Ca2+水平上升(P<0.05);缺氧可使MPTP开放度增加、?Ψm减低(P<0.05),且此时细胞内STC1、HIF-1α的基因表达升高(P<0.05);以上变化均随着缺氧程度的加剧而逐渐增大。结论缺氧条件下STC1、HIF-1α对Ca2+的负调控可能有利于肾癌细胞线粒体膜电势稳定,对肾癌细胞起保护作用。

Objective Detecting the change of mitochondrial membrane potential and stanniocalcin 1 （STC1） in renal carcinoma cell under hypoxic conditions, combined with cell proliferation and Ca2＋ levels, this preliminary study aim to explore the possible mechanisms of anti-hypoxia in renal cancer cells. Methods COC12 at the concentrations of 50 μ mol/L, 100 μ mol/L, 150μmol/L were added to the mediums of renal cancer ceils, and cells proliferation, expressions of HIF-1α, STC1, levels of Ca2＋, mitochondrial membrane potential and mitochondrial permeability transition pore （MPTP） were detected by MTT, RT-PCR, fluorescence spectrophotometer and ultraviolet spectrophotometer, respectively. Results Compared with the control group, hypoxia could inhibit cell proliferation significantly and increase intracellular Ca2＋ levels （P〈0.05）. Hypoxia could increase mitochondrial MPTP opening, reduce △ψm （P〈0.05）, and increase the expressions of HIF-1α, STC1 （P〈0.05）. The above changes were intensified with the degree of hypoxia increases gradually. Conclusion Ca2＋ negatively regulated by STC1, HIF-1α may be beneficial renal carcinoma stable mitochondrial membrane potential under hypoxic conditions.