槲皮素抑制HSP70表达的体外研究

Study on Inhibition of Heat Shock Protein 70 in vitro by Quercetin

目的探讨不同浓度槲皮素抑制正常细胞和肿瘤细胞热休克蛋白70(HSP70)的表达规律,寻找槲皮素抑制HSP70表达的最佳浓度,为研究HSP70的功能提供一种可行的方法。方法加入槲皮素至终浓度为50,100,150,200μmol/L,继续37℃培养6 h后,置42℃水浴受热1 h,于37℃恢复培养2 h,收获细胞。设立37℃培养组(正常对照)、42℃培养组(阳性对照)和0.1%DMSO组(溶剂对照)。分别处理肿瘤细胞株A549细胞和正常细胞株2BS细胞,用Western-blot检测细胞HSP70的表达水平。结果在A549细胞和2BS细胞中,随着槲皮素浓度的增加,HSP70表达逐渐降低。与正常对照组相比,阳性对照组和溶剂对照组的HSP70水平表达增高,差异有统计学意义(P<0.01)。在A549细胞中,当槲皮素浓度达到50μmol/L时,HSP70表达开始下调,但与阳性对照组相比,差异没有统计学意义(P>0.05);当槲皮素浓度达到100μmol/L时,与阳性对照组相比,HSP70表达显著降低,差异有统计学意义(P<0.05);当槲皮素浓度达到150,200μmol/L时,与阳性对照组相比,A549细胞HSP70表达降低更加明显(P<0.01);150μmol/L组和200μmol/L组相比,HSP70水平差异无统计学意义(P>0.05)。在2BS细胞中,当槲皮素浓度达到50μmol/L时,HSP70表达开始下调,与阳性对照组相比,差异有统计学意义(P<0.05);当槲皮素浓度达到100,150,200μmol/L时,与阳性对照组相比,HSP70水平也有显著性降低(P<0.01);150μmol/L组和200μmol/L组相比,HSP70水平差异无统计学意义(P>0.05)。结论槲皮素可以抑制肿瘤细胞和正常细胞HSP70的表达,150μmol/L的槲皮素处理A549和2BS细胞,抑制HSP70表达的效果最佳。

Objective To study the HSP70 expression in tumor cells and normal cells treated by different concentrations of quercetin. Methods Treatments of 50, 100, 150, 200 μmol/L quercetin were performed to A549 cells and 2BS cells at 37 ℃ for 6 h, then at 42 ℃ water bath for 1 h. Cells were cultured at 37℃C for 2 h recovery. Normal cultured cells were as a normal control. Cells treated by 42 ℃ were as a positive control and cells treated by 0. 1% DMSO were as a solvent control. HSP70 levels were detected by western blot assay. Results Along with the increasing quercetin concentrations from 50, 100, 150 to 200 μmol/L, HSP70 levels decreased. Compared with the group of normal control, there was a significant increase in the group of positive control or solvent control （P〈0.01）. There was no obvious difference between positive control and solvent control （P 〉 0.05）. For A549 cells, HSP70 levels decreased significantly in the group of 100, 150 and 200 μmol/L（P〈0.05 ,or P〈0.01）compared with positive control. There was no significant difference between 150 μmol/L and 200 μmol/L groups （P〉 0. 05）. For 2BS cells, compared with positive control, HSP70 levels decreased obviously in the group of 50 μmol/L （P（0.05）and in the groups of 100, 150, and 200μmol/L（P（0.01）, but there was no obvious difference between 150 μmol/L and 200 μmol/L groups（P）0.05）. Conclusions HSP70 expression can be inhibited in A549 or 2BS cells by quercetin, and the effect is the best with 150 μmol/L quercetin treatment.