Elia G, Santoro M G
Institute of Experimental Medicine, CNR, Rome, Italy.
Biochem J. 1994 May 15;300 ( Pt 1)(Pt 1):201-9. doi: 10.1042/bj3000201.
Synthesis of heat-shock proteins (HSPs) is universally induced in eukaryotic and prokaryotic cells by exposure to elevated temperatures or to other types of environmental stress. In mammalian cells, HSPs belonging to the 70 kDa family (HSP70) have a regulatory role in several cellular processes, and have been shown to be involved in the control of cell proliferation and differentiation. Although many types of HSP70 inducers have been identified, only a few compounds, all belonging to the flavonoid group, have been shown to inhibit HSP70 induction. Because inhibitors of HSP70 synthesis could be an important tool with which to study the function of this protein, we have investigated the effect of quercetin, a flavonoid with antiproliferative activity which is widely distributed in nature, on HSP70 synthesis in human K562 erythroleukaemia cells after treatment with severe or mild heat shock and with other inducers. Quercetin was found to affect HSP70 synthesis at more than one level, depending on the conditions used. Indeed, after severe heat shock (45 degrees C for 20 min) treatment with quercetin, at non-toxic concentrations, was found to inhibit HSP70 synthesis for a period of 3-4 h. This block appeared to be exerted at the post-transcriptional level and to be cell-mediated, as the addition of quercetin during translation of HSP70 mRNA in vitro had no effect. After prolonged (90 min) exposure at 43 degrees C, however, quercetin was found to inhibit also HSP70 mRNA transcription. Pretreatment of K562 cells with quercetin had no effect on HSP70 expression, and quercetin needed to be present during induction to be effective. Under all conditions tested, the quercetin-induced block of HSP70 synthesis was found to be transient and, after an initial delay, synthesis of HSP70 reached the control rate and continued at the same level for several hours after the time at which HSP70 synthesis had been turned off in control cells. Finally, inhibition of HSP70 synthesis by quercetin appeared to be dependent on the temperature used and on the type of stressor.
在真核细胞和原核细胞中,暴露于高温或其他类型的环境应激下会普遍诱导热休克蛋白(HSPs)的合成。在哺乳动物细胞中,属于70 kDa家族的热休克蛋白(HSP70)在多个细胞过程中发挥调节作用,并且已被证明参与细胞增殖和分化的控制。尽管已经鉴定出许多类型的HSP70诱导剂,但只有少数几种化合物(均属于黄酮类)被证明可抑制HSP70的诱导。由于HSP70合成抑制剂可能是研究该蛋白功能的重要工具,因此我们研究了槲皮素(一种具有抗增殖活性且在自然界中广泛分布的黄酮类化合物)对人K562红白血病细胞在经受重度或轻度热休克以及其他诱导剂处理后HSP70合成的影响。发现槲皮素根据所使用的条件在多个水平上影响HSP70的合成。实际上,在重度热休克(45℃处理20分钟)后,发现用无毒浓度的槲皮素处理可在3 - 4小时内抑制HSP70的合成。这种阻断似乎是在转录后水平发挥作用且由细胞介导,因为在体外HSP70 mRNA翻译过程中添加槲皮素没有效果。然而,在43℃长时间(90分钟)暴露后,发现槲皮素也会抑制HSP70 mRNA的转录。用槲皮素预处理K562细胞对HSP70的表达没有影响,并且槲皮素需要在诱导过程中存在才有效。在所有测试条件下,发现槲皮素诱导的HSP70合成阻断是短暂的,并且在初始延迟后,HSP70的合成达到对照速率,并在对照细胞中HSP70合成关闭后的数小时内持续保持在相同水平。最后,槲皮素对HSP70合成的抑制似乎取决于所使用的温度和应激源的类型。