三白脂素-8抑制缺氧诱导因子-1α对肾癌细胞解偶联功能的影响

Effect of Man A for inhibiting HIF-1α on uncoupling function of renal carcinoma cells

目的观察三白脂素-8(Man A)抑制缺氧诱导因子1α(HIF-1α)后对肾癌细胞的生长是否有抑制作用,此作用是否与线粒体解偶联功能相关。方法构建高表达HIF-1α的肾癌细胞模型,使用不同浓度Man A干预转染后的肾癌细胞(转染组)和单纯肾癌细胞(非转染组)。MTT法检测细胞增殖;RT-PCR及ELISA法检测细胞内HIF-1α、UCP2的表达;荧光分光光度计检测细胞内ATP水平;提取细胞线粒体,分光光度计检测细胞线粒体mPTP、ΔΨm的变化情况。结果 Man A干预细胞后,细胞内HIF-1α、UCP2的表达,ATP水平在高剂量组显著下降(P<0.05)。Man A抑制了肾癌细胞的增殖,且在中、高剂量出现显著下降(P<0.05)。Man A可提高肾癌细胞线粒体mPTP开放程度,并导致转染组所有剂量组,非转染组中高剂量组ΔΨm降低(P<0.05)。结论 Man A可显著抑制肾癌细胞增殖,并影响其线粒体解偶联功能。

Objective To observe whether manassantin A(Man A)inhibiting HIF-1αhaving the inhibiting effect on renal carcinoma cells growth and whether this effect being related with the uncoupling function of mitochondria.Methods The renal cancer cell models with HIF-1αhigh expression was constructed.Different concentrations of Man A solution were used to intervene the transfected renal cancer cell(transfection group)and simple renal cancer cells(non-transfection group).Then the cellular proliferation,expressions of HIF-1α,UCP2,mPTP and ΔΨm of cell mitochondria and levels of ATP were detected by MTT,RT-PCR,ELISA and fluorescence spectrophotometer respectively.Results After Man A intervening the cells,intracellular HIF-1α and UCP2 expressions,ATP level in the high dose group were decreased significantly(P<0.05)Man A inhibited the proliferation of renal carcinoma cells,moreover which in middle and low dosage groups were significantly decreased(P<0.05).Man A could increase the mitochondria mPTP open degree of renal cancer cells,and caused the ΔΨm decrease in all doses groups of transfection group and the middle and high dose group of the non-transfection group(P<0.05).Conclusion Man A can significantly inhibit the proliferation of renal carcinoma cells,and influences the uncoupling function of mitochondria.