低剂量氚水基于Nrf2通路诱导肺癌细胞的辐射适应性反应

摘要/Abstract

摘要： 本文研究了氚水在人非小细胞肺癌细胞A549中诱导辐射适应性反应及其高活度阈值,并进一步探究了Nrf2通路在氚水诱导适应性反应中的作用。设置7×10³、1.4×10⁴、2.1×10⁴、2.8×10⁴、3.5×10⁴ Bq/mL五个梯度氚水活度(累计照射剂量为0.55~8.27 mGy),用氚水预处理A549 细胞24 h,随后照射4 Gy X射线,用MTT检测细胞活力,显示较低活度(7×10³和1.4×10⁴ Bq/mL)氚水处理后可恢复4 Gy照射后A549细胞的细胞活力,具有适应性反应;而大于2.1×10⁴ Bq/mL活度的氚水处理后,A549的细胞活力不再恢复,适应性反应消失,随着活度的增加,细胞活力进一步降低;鉴于1.4×10⁴ Bq/mL的氚水诱导的适应性反应最为显著,因此,选用此活度氚水开展氧化水平的检测实验。结果显示,氚水可以显著降低X射线高剂量照射后胞内活性氧(ROS)水平,表明适应性反应受控于氧化水平;在氚水(1.4×10⁴ Bq/mL)预处理细胞阶段加入Nrf2的抑制剂(ML385),可显著抑制氚水诱导的辐射抗性,表明抗氧化途径Nrf2参与了此过程。

关键词: 氚水, 电离辐射, 适应性反应, Nrf2通路

Abstract: This study investigated the induction of radiation adaptive response and its high activity threshold by tritiated water (HTO) in human non-small cell lung cancer cells (A549). The role of the Nrf2 pathway in the induction of adaptive response by HTO was further explored. Five gradient activities of HTO (7×10³, 1.4×10⁴, 2.1×10⁴, 2.8×10⁴, 3.5×10⁴ Bq/mL) were set (accumulated radiation dose ranging from 0.55 to 8.27 mGy). A549 cells were pre-treated with HTO for 24 hours, followed by irradiation with 4 Gy X-rays. Cell viability was measured using MTT assay. The results showed that lower activities of HTO (7×10³ and 1.4×10⁴ Bq/mL) could restore the cell viability of A549 cells after 4 Gy irradiation, indicating the presence of an adaptive response. However, HTO with activities higher than 2.1×10⁴ Bq/mL resulted in no recovery of cell viability in A549 cells, indicating the disappearance of adaptive response. Moreover, with increasing activity, cell viability further decreased. Considering that the adaptive response induced by HTO with an activity of 1.4×10⁴ Bq/mL is the most significant, this activity of HTO was selected for the detection of oxidative levels. The results showed that HTO can significantly reduce the intra-cellular reactive oxygen species (ROS) levels after high-dose X-ray irradiation, indicating that the adaptive response is regulated by oxidative levels. Moreover, the addition of Nrf2 inhibitor (ML385) during the pre-treatment stage of HTO (1.4×10⁴ Bq/mL) can significantly inhibit the radiation resistance induced by HTO, suggesting the involvement of the antioxidant pathway Nrf2 in this process.

Key words: HTO, ionizing radiation, adaptive response, Nrf2 pathway

中图分类号:

Q691

丁子仪, 徐照, 曹敬文, 季涛涛, 周袁媛, 杨彦彦, 李桃生. 低剂量氚水基于Nrf2通路诱导肺癌细胞的辐射适应性反应[J]. 辐射防护, 2025, 45(1): 67-76.

DING Ziyi, XU Zhao, CAO Jingwen, JI Taotao, ZHOU Yuanyuan, YANG Yanyan, LI Taosheng. Low dose tritiated water induced radiation-adaptive responses in lung cancer cells based on the Nrf2 pathway[J]. RADIATION PROTECTION, 2025, 45(1): 67-76.

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