【摘要】 目的:研究缺氧条件下斯钙素蛋白1(STC-1)下调钙离子及缺氧诱导因子1α(HIF-1α)水平后对肾癌细胞增殖平衡的影响。方法:构建正常及缺氧两种条件肾癌细胞(GRC-1)模型,分别用0.1、0.5、1.0 nmol/L的STC-1溶液干预48h,并设空白对照(只加生理盐水)。MTT法检测肾癌细胞生长情况,逆转录PCR和ELISA方法检测细胞内STC-1、HIF-1α的基因和蛋白表达,荧光分光光度计检测细胞内钙离子水平,分光光度计检测三磷酸腺苷(ATP)含量。结果:缺氧可使肾癌细胞内HIF-1α、STC-1基因和蛋白的表达和钙离子水平均显著升高(均P<0.05),而外源性STC-1可逆转上述改变(均P<0.05),并存在量效关系;缺氧明显抑制肾癌细胞的生长和ATP的生成(均P<0.05),而外源性STC-1亦可逆转上述改变(均P<0.05),随着外源性STC-1浓度的增高,ATP生成逐渐增加,但STC-1对两种条件下肾癌细胞模型的增殖促进作用却减少。结论:外源性STC-1可能通过下调细胞内钙离子含量,提高ATP产量来促进肾癌细胞的抗缺氧增殖,但对HIF-1α的进行性抑制又阻碍了肾癌细胞的增殖,该作用可能参与了肾癌抗缺氧增殖的机制。更多还原

【Abstract】 Objective:To investigate effects of stanniocalcin-1（STC-1） on proliferation balance under hypoxic condition in renal cancer cells and its mechanism.Methods:Hypoxic model was induced on renal cancer GRC-1 cells（Group H）,the cells were treated with STC-1 protein at concentrations of 0.1 nmol/L（HI）,0.5nmol/L（H2）,1.0 nmol/L（H3）,or normal saline（HO） for 48 h,respectively.Cells proliferation was measured by MTT assay:mRNA and protein expressions of hypoxia inducible factor la（HIF-1α） and STC-1 in GRC-1 cells were detected by RTPCR and ELISA,respectively:the intracellular levels of Ca²⁺ and adenosine triphosphate（ATP） were determined by fluorescence spectrophotometry and spectrophotometry,respectively.Results:The expression of HIF-1α,STC-1 and Ca²⁺levels were increased in GRC-1 cells under hypoxia condition:STC-1 reversed these changes in a dose-effect manner.Hypoxia significantly inhibited cell proliferation and the generation of ATP in GRC-1 cells and exogenous STC-1 reversed the effects of hypoxia:ATP generation increased gradually with increasing STC-1 concentration,but the cell proliferation was reduced.Conclusion:Exogenous STC-1 can promote the proliferation of renal cancer cells in hypoxia condition by reducing HIF-la expression and Ca²⁺ content and increased ATP production,but the progressive inhibition of HIF-1α hindered the renal carcinoma cell proliferation further,which indicates that STC-1may be involved in anti-hypoxia proliferative balance of renal cancer cells.更多还原