摘要
目的探讨索拉菲尼对肾癌细胞抑制效应与斯钙素蛋白1(STC-1)调控细胞内钙稳态之间的关系,以期寻找逆转肾癌细胞耐药性的新途径。方法分别以0、1.25、2.5、5、10、20、40μmol/L索拉非尼干预肾癌细胞,按浓度依次递增进行分组,MTT法检测肾癌细胞增殖情况;RT-PCR方法及ELLSA法检测各组细胞STC-1的基因及蛋白表达;荧光分光光度法检测各组细胞内钙离子含量;化学比色法测定各组细胞Ca^(2+)-ATP酶活性。结果随着索拉菲尼浓度的递增,出现10μmol/L的浓度转折点,细胞抑制率及Ca^(2+)含量由逐渐升高趋向平稳,Ca^(2+)-ATP酶活性由逐渐下降趋向平稳,STC-1基因及蛋白表达逐渐升高。结论STC-1可能是肾癌细胞恶性增殖的保护因素,其可能通过调控Ca^(2+)水平,降低Ca^(2+)-ATP酶活性,使肾癌细胞内线粒体能量代谢活跃,促进肾癌细胞适应缺氧环境,从而对索拉菲尼产生抵抗作用。
Objective To explore the relationship between the inhibitory effect of sorafenib on renal cell carcinoma cells and the regulation of intracellular calcium homeostasis by STC-1,so as to find a new way to reverse drug resistance of renal cell carcinoma cells.Methods Sorafenib was used to intervene renal cancer cells at concentrations of 0,1.25,2.5,5,10,20 and 40μmol/L,and groups were grouped according to increasing concentrations.The proliferation of renal cancer cells was detected by MTT method.RT-PCR method and ELLSA method was used to detect the gene and protein expression of STC-1 in each group of cells.Fluorescence spectrophotometry was used to detect the calcium ion content in each group of cells;chemical colorimetry was used to determine the activity of Ca^(2+)-ATPase in each group of cells.Results With the increasing concentration of sorafenib,a concentration turning point of 10μmol/L appeared.The cell inhibition rate and Ca^(2+)content gradually increased to stabilize,the Ca^(2+)-ATPase activity gradually decreased to stabilize,and the STC-1 gene and Protein expression gradually increased.Conclusion STC-1 may be a protective factor for the malignant proliferation of renal cancer cells.STC-1 may regulate the level of Ca^(2+),reduce the activity of Ca^(2+)-ATPase,activate the energy metabolism of mitochondria in renal cancer cells,and promote the adaptation of renal cancer cells to hypoxia,so as to produce resistance to Solafeni.
作者
沈亚楠
谷江
张永春
罗浩鸣
Shen Yanan;Gu Jiang;Zhang Yongchun(The Affiliated Hospital of Guizhou Medical University,Guizhou 550000,China)
出处
《医学研究杂志》
2021年第6期98-101,97,共5页
Journal of Medical Research
基金
贵州省科技厅社会攻关计划项目[黔科合sy字(2011)3060]。
