斯钙素蛋白1下调钙离子及缺氧诱导因子1α水平调控肾癌细胞抗缺氧增殖平衡  

STC-1 is involved in anti-hypoxia proliferative balance of renal cancer cells by down-regulation of intracellular Ca2+and HIF-1α levels

作　　者：朱致晖[1] 谷江[1] 张永春[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]扬州市江都人民医院泌尿外科,江苏扬州225200

出　　处：《浙江大学学报（医学版）》2014年第5期528-534,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：贵州省科技支撑计划

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.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 （H1）, 0.5 nmol/L （ H2 ） , 1.0 nmol/L （ H3 ）, or normal saline （ HO ） for 48 h, respectively. Cells proliferation was measured by MTF assay; mRNA and protein expressions of hypoxia inducible factor 1 α （HIF-1 α） and STC-1 in GRC-1 cells were detected by RT- PCR and ELISA, respectively; the intracellular levels of Ca^2＋ and adenosine triphosphate （ ATP ） were determined by fluorescence spectrophotometry and spectrophotometry, respectively. Results： The expression of HIF-1α, STC-1 and Ca^2＋ 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-1α expression and Ca^2＋ content and increased ATP production, but the progressive inhibition of HIF- 1α hindered the renal carcinoma cell proliferation further, which indicates that STC-1 may be involved in anti-hypoxia proliferative balance of renal cancer cells.

关 键 词：癌 肾细胞/病理学 缺氧 缺氧诱导因子1 Α亚基 钙 腺苷三磷酸 

分 类 号：R737.11[医药卫生—肿瘤]