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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 investigate the expression of heat shock protein 70 (HSP70) in normal cells and tumor cells with different concentrations of quercetin, and to find the optimal concentration of quercetin for inhibiting the expression of HSP70, and to provide a feasible method for studying the function of HSP70. Methods Quercetin was added to final concentrations of 50, 100, 150, and 200 μmol/L. After incubation at 37°C for 6 h, the cells were heated in a 42°C water bath for 1 h, and cultured at 37°C for 2 h to harvest cells. A 37°C culture group (normal control), a 42°C culture group (positive control), and a 0.1% DMSO group (solvent control) were set up. The tumor cell line A549 cells and normal cell line 2BS cells were treated respectively, and the expression level of HSP70 was detected by Western-blot. Results In A549 cells and 2BS cells, the expression of HSP70 gradually decreased as the concentration of quercetin increased. Compared with the normal control group, the expression levels of HSP70 in the positive control group and the solvent control group increased, the difference was statistically significant (P<0.01). In A549 cells, when quercetin concentration reached 50μmol/L, HSP70 expression began to decrease, but there was no statistically significant difference (P>0.05) compared with the positive control group; when the quercetin concentration reached 100μmol/L Compared with the positive control group, the expression of HSP70 was significantly decreased (P<0.05). When the concentration of quercetin reached 150 and 200 μmol/L, the expression of HSP70 in A549 cells decreased more significantly than that in the positive control group. (P<0.01); There was no significant difference in HSP70 level between 150μmol/L group and 200μmol/L group (P>0.05). In 2BS cells, when quercetin concentration reached 50μmol/L, HSP70 expression began to decrease, compared with the positive control group, the difference was statistically significant (P<0.05); when the quercetin concentration reached 100,150,200μmol/L Compared with the positive control group, the level of HSP70 was also significantly decreased (P<0.01). Compared with the 200 μmol/L group, there was no significant difference in the level of HSP70 between the 150 μmol/L group and the 200 μmol/L group (P>0.05). Conclusion Quercetin can inhibit the expression of HSP70 in tumor cells and normal cells. Treatment of A549 and 2BS cells with 150 μmol/L quercetin has the best effect in inhibiting the expression of HSP70.