Wang Qian, Zhang Qiuning, Wang Xiaohu, Luo Hongtao, Du Tianqi, Wu Luyao, Tan Mingyu, Chen Yanliang, Wu Xun, Sun Shilong, Liu Zhiqiang, Xie Yi, Yuan Wenzhen
The First School of Clinical Medicine, Lanzhou University, Lanzhou 730030, China.
Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730030, China.
Biomedicines. 2024 Sep 30;12(10):2231. doi: 10.3390/biomedicines12102231.
: Cancer cells with 'stemness' are generally resistant to chemoradiotherapy. This study aims to compare the differences in radiation sensitivity of A549 and CD44A549 stem-like cells to X-rays and carbon ion radiation (C-ions), and to find a target that can kill cancer stem-like cells (CSCs) of non-small cell lung cancer (NSCLC). : The study used two cell lines (A549 and CD44A549). The tumorigenicity of cells was tested with animal experiments. The cells were irradiated with X-rays and C-ions. Cell viability was detected using the CCK-8 and EdU assay. A liquid chromatograph-mass spectrometer (LC-MS) helped detect metabolic differences. Protein and mRNA expression were detected using a Western blot, reverse transcription-quantitative (RT-qPCR), and PCR array. The autophagic activity was monitored with a CYTO-ID Autophagy Detection Kit 2.0. Immunofluorescence and co-immunoprecipitation helped to observe the localization and interaction relationships. First, we verified the radio-resistance of CD44A549 stem-like cells. LC-MS indicated the difference in autophagy between the two cells, followed by establishing a correlation between the radio-resistance and autophagy. Subsequently, the PCR array proved that TGM2 is significantly upregulated in CD44A549 stem-like cells. Moreover, the TGM2 knockdown by small interfering RNA could decrease the radio-resistance of CD44A549 cells. Bioinformatic analyses and experiments showed that TGM2 is correlated with the expression of CD44 and LC3B. Additionally, TGM2 could directly interact with LC3B. We established the CD44-TGM2-LC3 axis: CD44 mediates radio-resistance of CD44A549 stem-like cells through TGM2 regulation of autophagy. Our study may provide new biomarkers and strategies to alleviate the radio-resistance of CSCs in NSCLC.
具有“干性”的癌细胞通常对放化疗具有抗性。本研究旨在比较A549和CD44A549干细胞样细胞对X射线和碳离子辐射（C离子）的辐射敏感性差异，并寻找能够杀死非小细胞肺癌（NSCLC）癌干细胞样细胞（CSCs）的靶点。：该研究使用了两种细胞系（A549和CD44A549）。通过动物实验检测细胞的致瘤性。用X射线和C离子对细胞进行照射。使用CCK-8和EdU检测法检测细胞活力。液相色谱-质谱联用仪（LC-MS）有助于检测代谢差异。使用蛋白质免疫印迹法、逆转录定量（RT-qPCR）和PCR阵列检测蛋白质和mRNA表达。使用CYTO-ID自噬检测试剂盒2.0监测自噬活性。免疫荧光和免疫共沉淀有助于观察定位和相互作用关系。首先，我们验证了CD44A549干细胞样细胞的抗辐射性。LC-MS显示了两种细胞之间自噬的差异，随后建立了抗辐射性与自噬之间的相关性。随后，PCR阵列证明TGM2在CD44A549干细胞样细胞中显著上调。此外，小干扰RNA敲低TGM2可降低CD44A549细胞的抗辐射性。生物信息学分析和实验表明，TGM2与CD44和LC3B的表达相关。此外，TGM2可直接与LC3B相互作用。我们建立了CD44-TGM2-LC3轴：CD44通过TGM2调节自噬介导CD44A549干细胞样细胞的抗辐射性。我们的研究可能为减轻NSCLC中CSCs的抗辐射性提供新的生物标志物和策略。
