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作 者:谷江[1] 张永春[1] 朱致晖[1] 杨清滔[1] 杨永安[1] 王楠[1] 祝庆亮[2]
机构地区:[1]贵阳医学院附属医院泌尿外科,贵州贵阳550004 [2]江都人民医院泌尿外科,江苏扬州225200
出 处:《分子诊断与治疗杂志》2014年第1期42-46,共5页Journal of Molecular Diagnostics and Therapy
基 金:贵州省科技厅社会攻关计划项目(黔科合SY字[2011]3060)
摘 要: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.
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