PO.MCB10.02 · 分子与细胞生物学
piRNA-5939通过PIWIL3介导的转录后调控驱动肺癌进展
piRNA-5939 drives lung cancer progression via PIWIL3-mediated post-transcriptional regulation
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：PIWI相互作用RNA（piRNA）是一类独特的小非编码RNA，最初因其在生殖系转座子沉默中的作用而被描述。近期证据提示体细胞piRNA参与癌症相关基因调控；然而，其对肺肿瘤进展的机制性贡献仍不明确。在本研究中，我们鉴定piRNA-5939为一种新的致癌调控因子，其通过PIWIL3介导的基因表达转录后调节增强肺癌细胞增殖和迁移。
方法：将人肺腺癌细胞系（A549、ABC1和H522）转染针对piRNA-5939的反义寡核苷酸。通过MTT、Click-iT™ EdU掺入和集落形成试验评估增殖；通过划痕愈合和Transwell试验分析迁移和侵袭；使用Annexin V Alexa Fluor™ 488/碘化丙啶流式细胞术定量凋亡。通过RNA测序检查piRNA-5939敲低后的整体转录组变化，并使用qRT-PCR和Western blot验证关键靶点。细胞质和细胞核RNA分离测定piRNA-5939的亚细胞定位及其对PIWIL家族（PIWIL1-4）表达谱的影响。
结果：亚细胞分离揭示piRNA-5939在各肺癌细胞系中呈异质性的细胞质和细胞核分布。基线分析表明PIWIL3转录本优先定位于细胞质，而piRNA-5939沉默导致PIWIL3 mRNA和蛋白表达显著下调。在功能上，piRNA-5939缺失相对于对照显著降低细胞增殖（约40%）和迁移（约55%）（p < 0.01）。转录组分析突显了致癌信号通路的抑制，包括调控细胞周期进展、细胞骨架重塑和RNA稳定性的通路——均与PIWIL3活性减弱一致。
结论：这些发现将piRNA-5939鉴定为一种功能性致癌piRNA，其通过PIWIL3依赖性的关键生长和迁移通路转录后调控促进肺癌进展。piRNA-5939/PIWIL3轴代表了一种此前未被认识的调控机制以及肺癌中潜在的治疗脆弱点。
查看英文原文 English abstract
Introduction: PIWI-interacting RNAs (piRNAs) constitute a distinct class of small non-coding RNAs initially described for their roles in germline transposon silencing. Recent evidence implicates somatic piRNAs in cancer-associated gene regulation; however, their mechanistic contribution to lung tumor progression remains poorly defined. In this study, we identify piRNA-5939 as a novel oncogenic regulator that enhances lung cancer cell proliferation and migration via PIWIL3-mediated post-transcriptional modulation of gene expression.
Methods: Human lung adenocarcinoma cell lines (A549, ABC1, and H522) were transfected with antisense oligonucleotides targeting piRNA-5939. Proliferation was assessed by MTT, Click-iT™ EdU incorporation, and colony formation assays; migration and invasion were analyzed by wound-healing and transwell assays; and apoptosis was quantified using Annexin V Alexa Fluor™ 488/Propidium Iodide flow cytometry. Global transcriptomic changes following piRNA-5939 knockdown were examined by RNA sequencing, and key targets were validated using qRT-PCR and Western blotting. Cytoplasmic and nuclear RNA fractionation determined piRNA-5939 subcellular localization and its impact on PIWIL family (PIWIL1-4) expression profiles.
Results: Subcellular fractionation revealed heterogeneous cytoplasmic and nuclear distribution of piRNA-5939 across lung cancer cell lines. Baseline profiling demonstrated that PIWIL3 transcripts are preferentially localized to the cytoplasm, and piRNA-5939 silencing led to a significant downregulation of PIWIL3 mRNA and protein expression. Functionally, loss of piRNA-5939 markedly reduced cell proliferation (~40%) and migration (~55%) relative to controls (p < 0.01). Transcriptomic analyses highlighted suppression of oncogenic signaling pathways, including those governing cell cycle progression, cytoskeletal remodeling, and RNA stability-all consistent with attenuated PIWIL3 activity.
Conclusions: These findings identify piRNA-5939 as a functional oncogenic piRNA that promotes lung cancer progression through PIWIL3-dependent post-transcriptional regulation of key growth and migration pathways. The piRNA-5939/PIWIL3 axis represents a previously unrecognized regulatory mechanism and a potential therapeutic vulnerability in lung cancer.
利益披露 Disclosure
S. K. Rai, None..
Y. Deng, None.