Zhong Yazhen, Lin Zechen, Lu Jinhua, Lin Xianlei, Xu Wei, Wang Nan, Huang Siyu, Wang Yuanyuan, Zhu Yuan, Chen Zhu, Lin Shengyou
Oncology Department, Hangzhou Hospital of Traditional Chinese Medicine, GuangXing Hospital Affiliated to Zhejiang Chinese Medical University Hangzhou, China.
Fourth Clinical Medical College, Hangzhou Cancer Hospital Hangzhou, China.
Am J Transl Res. 2019 Feb 15;11(2):733-743. eCollection 2019.
Radiation-induced lung toxicity, including radiation pneumonitis and pulmonary fibrosis, often occurs in patients receiving radiation therapy. Epithelial-mesenchymal transition (EMT) of alveolar epithelial cells (AECs) plays critical roles in radiation-induced lung toxicity. In the present study, RNA sequencing was applied to examine the whole transcriptomes of human pulmonary AEC cells (HPAEpiC) with or without radiation treatment. We found that cytokine, chemokine and cell adhesion signaling pathways were enriched in radiation-treated cells. CCL2 (C-C Motif Chemokine Ligand 2), CCL5 and CCR4 (C-C Motif Chemokine Receptor 4) were among the top enriched genes in chemokine signaling pathway. The upregulation of CCL2, CCL5 and CCR4 in response to irradiation was confirmed at both mRNA and protein levels by real-time PCR, western blotting and enzyme-linked immunosorbent assay analyses. Ophiopogonin B, a bioactive ingredient of Radix , was found to attenuate radiation-induced EMT in HPAEpiC cells as demonstrated by the alteration in cell morphology, and the expression of E-cadherin and Vimentin. Ophiopogonin B could also reduce radiation-induced expression of CCL2, CCL5, CCR4 and phosphorylated ERK (p-ERK). Moreover, CCR4 knockdown, U0126 (a MEK/ERK inhibitor) or ophiopogonin B also partially blocked the EMT promoting effects of CCL2 and CCL5. Our data suggested CCL2, CCL5 and CCR4 may be potential therapeutic targets for radiation-induced lung toxicity. Ophiopogonin B, which could down-regulate CCL2, CCL5 and CCR4, may be a useful radioprotective agent.
辐射诱导的肺毒性，包括放射性肺炎和肺纤维化，经常发生在接受放射治疗的患者中。肺泡上皮细胞（AECs）的上皮-间质转化（EMT）在辐射诱导的肺毒性中起关键作用。在本研究中，应用RNA测序来检测接受或未接受辐射治疗的人肺AEC细胞（HPAEpiC）的整个转录组。我们发现细胞因子、趋化因子和细胞黏附信号通路在接受辐射处理的细胞中富集。CCL2（C-C基序趋化因子配体2）、CCL5和CCR4（C-C基序趋化因子受体4）是趋化因子信号通路中富集程度最高的基因之一。通过实时PCR、蛋白质印迹和酶联免疫吸附测定分析，在mRNA和蛋白质水平均证实了CCL2、CCL5和CCR4在辐射响应中的上调。麦冬皂苷B是麦冬的一种生物活性成分，通过细胞形态的改变以及E-钙黏蛋白和波形蛋白的表达，发现其可减轻辐射诱导的HPAEpiC细胞中的EMT。麦冬皂苷B还可降低辐射诱导的CCL2、CCL5、CCR4和磷酸化ERK（p-ERK）的表达。此外，CCR4敲低、U0126（一种MEK/ERK抑制剂）或麦冬皂苷B也部分阻断了CCL2和CCL5的EMT促进作用。我们的数据表明CCL2、CCL5和CCR4可能是辐射诱导的肺毒性的潜在治疗靶点。可下调CCL2、CCL5和CCR4的麦冬皂苷B可能是一种有用的辐射防护剂。
