Graduate School of Pharmaceutical Sciences, Ewha Women's University, Seoul, Korea.
Division of Radiation Effects, Korea Institute of Radiological and Medical Sciences, Seoul, Korea.
Int J Radiat Oncol Biol Phys. 2015 Mar 15;91(4):807-16. doi: 10.1016/j.ijrobp.2014.11.031.
We previously screened natural compounds and found that coniferyl aldehyde (CA) was identified as an inducer of HSF1. In this study, we further examined the protective effects of CA against ionizing radiation (IR) in normal cell system.
Western blotting and reverse transcription-polymerase chain reaction tests were performed to evaluate expression of HSF1, HSP27, and HSP70 in response to CA. Cell death and cleavage of PARP and caspase-3 were analyzed to determine the protective effects of CA in the presence of IR or taxol. The protective effects of CA were also evaluated using animal models.
CA increased stability of the HSF1 protein by phosphorylation at Ser326, which was accompanied by increased expression of HSP27 and HSP70. HSF1 phosphorylation at Ser326 by CA was mediated by EKR1/2 activation. Cotreatment of CA with IR or taxol in normal cells induced protective effects with phosphorylation- dependent patterns at Ser326 of HSF1. The decrease in bone marrow (BM) cellularity and increase of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive BM cells by IR were also significantly inhibited by CA in mice (30.6% and 56.0%, respectively). A549 lung orthotopic lung tumor model indicated that CA did not affect the IR-mediated reduction of lung tumor nodules, whereas CA protected normal lung tissues from the therapeutic irradiation.
These results suggest that CA may be useful for inducing HSF1 to protect against normal cell damage after IR or chemotherapeutic agents.
我们之前筛选了天然化合物，发现松柏醛（CA）可被鉴定为 HSF1 的诱导物。在本研究中，我们进一步研究了 CA 对正常细胞系统中电离辐射（IR）的保护作用。
通过 Western blot 和逆转录-聚合酶链反应试验评估了 HSF1、HSP27 和 HSP70 的表达，以评估 CA 的响应。通过分析细胞死亡和 PARP 和 caspase-3 的裂解来确定 CA 在存在 IR 或紫杉醇时的保护作用。还使用动物模型评估了 CA 的保护作用。
CA 通过 Ser326 磷酸化增加了 HSF1 蛋白的稳定性，同时增加了 HSP27 和 HSP70 的表达。CA 诱导的 HSF1 的 Ser326 磷酸化是通过 EKR1/2 激活介导的。在正常细胞中，CA 与 IR 或紫杉醇共同处理诱导了具有 HSF1 Ser326 磷酸化依赖性的保护作用。IR 引起的骨髓（BM）细胞减少和末端脱氧核苷酸转移酶 dUTP 缺口末端标记阳性 BM 细胞增加也被 CA 在小鼠中显著抑制（分别为 30.6%和 56.0%）。A549 肺原位肺肿瘤模型表明，CA 不影响 IR 介导的肺肿瘤结节减少，但 CA 保护正常肺组织免受治疗性照射。
这些结果表明，CA 可能有助于诱导 HSF1 以保护正常细胞免受 IR 或化疗药物的损伤。
