不同程度放射性肺炎非小细胞肺癌患者血浆外泌体microRNA表达差异分析

Differential Expression of Plasma Exosome microRNAs in Non-small Cell Lung Cancer Patients with Different Degree of Radiation Pneumonia

[目的]分析发生不同程度放射性肺炎(radiation pneumonia,RP)的Ⅲ期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的血浆外泌体microRNA(miRNA)表达差异谱,并应用生物信息学分析差异miRNAs的作用。[方法]收集NSCLC患者放疗前血浆,根据放疗完成后发生放射性肺炎程度分为RP≤1级组和RP>1级组,从中筛选出5对患者。分离血浆中的外泌体,通过二代测序技术对miRNAs进行精准的定量,并进行差异分析。对差异miRNAs进行靶基因预测注释,并进行生物信息学分析,分析其生物学功能及信号通路。[结果]与RP>1级的患者相比,RP≤1级患者血浆外泌体中15个miRNAs有显著表达差异(Log2FC>3,P<0.05)。通过生物信息学分析发现,差异靶基因功能主要富集于TGF-β、Hippo、MAPK、mTOR等信号通路,涉及RNA聚合酶、泛素样蛋白转移酶等多种细胞生化代谢途径,并参与细胞衰老、细胞周期、细胞连接等多种生物学过程。[结论]发生不同程度放射性肺炎NSCLC患者放疗前血浆外泌体miRNA表达谱有明显差异,可能通过Hippo、TGF-β等信号通路在放射性肺炎的发生发展中发挥生物学调控作用。

[Objective]To analyze the differential expression profile of plasma exosome microRNA(miRNA) in stage Ⅲ non-small cell lung cancer(NSCLC) patients with different degrees of radiation pneumonia(RP). [Methods]The plasma of NSCLC patients before radiotherapy was collected. After radiotherapy the patients were divided into RP≤1 group and RP>1 group according to the degree of RP, from which 5 pairs were selected for further study. Exosomes in plasma were isolated, and exosome miRNAs were quantified by second-generation sequencing techniques, and differential expression profile of miRNAs between RP ≤1 group and RP >1 group was analyzed.Target genes were predicted and annotated for differential miRNAs, and bioinformatics analysis was conducted for their biological functions and signaling pathways. [Results]The expression of15 miRNAs in plasma exosomes of RP≤1 patients was significantly different from that of RP>1patients(Log2FC>3, P<0.05). Bioinformatic analysis revealed that the differential target genes were mainly enriched in TGF-β, Hippo, MAPK, mTOR and other signaling pathways, involved in a variety of cellular biochemical metabolic pathways such as RNA polymerase and ubiquitin like protein transferase, and involved in a variety of biological processes such as cell senescence, cell cycle and cell connection. [Conclusion]The miRNA expression profiles in plasma exosomes of NSCLC patients with different degrees of RP before radiotherapy are significantly different, which may be involved in the occurrence and development of RP through Hippo, TGF-β and other signaling pathways.