Chen Xu, Kong Rui, Qi Yaxian, Li Lingchen, Yin Chenrui, Sun Lingyou, Jian Chunli, Cai Ping, Yang Qiao, Sun Jianguo
Department of Oncology, Xinqiao Hospital, Army Medical University, Chongqing, China.
Department of Medical Affairs, Xinqiao Hospital, Army Medical University, Chongqing, China.
J Cancer Res Clin Oncol. 2025 Sep 18;151(10):262. doi: 10.1007/s00432-025-06310-x.
Overcoming miRNA-mediated radioresistance and enhancing its synergy with immunotherapy remained significant challenges.
A total of 23 patients with locally advanced non-small cell lung cancer (NSCLC) undergoing thoracic radiotherapy from a single center were enrolled. Pre-radiotherapy blood samples were collected and analyzed using real time qPCR array to detect miRNA expression profiles, identifying differential miRNAs between responders and non-responders. In vitro experiments further assessed the impact of radiotherapy on significant differential miRNAs. Targeted immune genes of miRNAs were predicted through bioinformatics websites and validated by cellular experiments. Using TCGA and GEO datasets, the association between immune gene of interest and survival outcomes and immune infiltration were investigated. In vivo experiment was further performed to investigate the relationship between dendritic cell (DC) expression and miR-26b-5p following radiotherapy.
Using pre-radiotherapy blood samples from 23 NSCLC patients, 22 differentially expressed miRNAs were identified between responders and non-responders. Among them, miR-26b-5p exhibited significant differential expression, suggesting its role as a potential radioresistant molecule. The dual-luciferase assay confirmed miR-26b-5p targeted PRKCD, an immune-related gene. After continuous three days of 2-Gy irradiation, the expression of miR-26b-5p decreased significantly, while the expression of PRKCD increased. The effect of radiotherapy on PRCKD expression were further validated in clinical samples, which demonstrated elevated PRCKD expression after thoracic radiotherapy. Bioinformatic analysis using TCGA and GEO datasets revealed that a higher PRKCD expression was correlated with better survival outcomes, increased immune cell infiltration, and better outcomes. Further in vivo experiments showed that, after radiotherapy, the inhibition of miR-26b-5p showed a significantly higher proportion of DCs than the controls, along with increased expression of CD80, CD86, and TNFSF4.
miR-26b-5p and PRKCD modulates dual resistance of both radiotherapy and immunotherapy in NSCLC. These insights demonstrate that downregulating miR-26b-5p could offer a promising therapeutic strategy to enhance radiosensitivity and immune responses.
克服微小RNA（miRNA）介导的放射抗性并增强其与免疫疗法的协同作用仍然是重大挑战。
纳入了来自单一中心的23例接受胸部放疗的局部晚期非小细胞肺癌（NSCLC）患者。采集放疗前血样并使用实时定量PCR阵列进行分析，以检测miRNA表达谱，确定应答者和非应答者之间的差异miRNA。体外实验进一步评估了放疗对显著差异miRNA的影响。通过生物信息学网站预测miRNA的靶向免疫基因，并通过细胞实验进行验证。使用TCGA和GEO数据集，研究了感兴趣的免疫基因与生存结果及免疫浸润之间的关联。进一步进行体内实验以研究放疗后树突状细胞（DC）表达与miR-26b-5p之间的关系。
使用23例NSCLC患者的放疗前血样，在应答者和非应答者之间鉴定出22种差异表达的miRNA。其中，miR-26b-5p表现出显著差异表达，表明其作为潜在放射抗性分子的作用。双荧光素酶测定证实miR-26b-5p靶向PRKCD，一个免疫相关基因。连续三天进行2 Gy照射后，miR-26b-5p的表达显著降低，而PRKCD的表达增加。放疗对PRCKD表达的影响在临床样本中得到进一步验证，结果显示胸部放疗后PRCKD表达升高。使用TCGA和GEO数据集进行的生物信息学分析表明，较高的PRKCD表达与更好的生存结果、增加的免疫细胞浸润和更好的结局相关。进一步的体内实验表明，放疗后，抑制miR-26b-5p组的DC比例显著高于对照组，同时CD80、CD86和TNFSF4的表达增加。
miR-26b-5p和PRKCD调节NSCLC中放疗和免疫疗法的双重抗性。这些见解表明，下调miR-26b-5p可能提供一种有前景的治疗策略，以增强放射敏感性和免疫反应。
