TGF-β-lncAF147375-Notch信号通路参与肿瘤微环境介导肺癌骨转移的机制研究

Lung cancer is the most common malignant tumor in terms of both incidence and mortality in China. About 1/3 of patients with lung cancer suffer from bone metastasis, which could cause severe bone pain, pathologic fracture and paraplegia, seriously impact on quality of life with lung cancer patients and bring trouble to clinical treatment. However, chemotherapy or novel molecular target drugs did not achieve satisficed efficacy in treatment with bone metastasis. Our group persists in developing the mechanism of lung cancer bone metastasis and our previous works showed TGF/Notch3 –EMT signal pathway involved in bone metastasis of lung cancer, in which the remodeling of lung cancer cells to bone microenvironment played a vital role in the complex process. Long non-coding RNAs (lncRNAs) are emerging as new players in the cancer paradigm demonstrating potential roles in both oncogenic and tumor suppressive pathways. To explore the key molecular involved in TGF/Notch-EMT signal pathway, our previous works firstly performed high-through lncRNAs screening based on both the TGF-induced EMT cell lines and bone metastasis tissues. A serious of loss and gain function assay was performed and lncRNA -AF147375 was indicated as a pivotal factor in the lung cancer cells interaction with bone microenvironment and subsequently induced bone metastasis, but detailed mechanism is not clear. In this study, series of molecular biological approaches will be employed to further investigate the role and exact mechanism of lncRNA -AF147375 and its candidates target genes in the remodeling of lung cancer cells to bone microenvironment. This work will help us understand more about the molecular mechanism of lung cancer bone metastasis and help us to explore new drug in the treatment of lung cancer bone metastasis.

肺癌是我国发病率和死亡率最高的恶性肿瘤，约1/3肺癌患者会发生骨转移而导致剧烈疼痛、骨折和截瘫，新一代化疗或靶向药物在治疗肺癌骨转移瘤中未获得满意疗效。申请者长期致力于肺癌骨转移机制研究，首次报道了TGF/Notch-EMT是介导肺癌细胞与骨微环境相互作用促进骨转移发生的关键通路。为了寻求该通路中的关键“分子”,申请者基于前期建立的TGF-EMT细胞模型及发生骨转移的肺癌组织进行lncRNA双重筛选，通过共表达分子网络分析、临床标本验证、体内外功能试验发现了一个参与肺癌骨转移的lncRNA新分子-lnc-AF147375，初步研究证实该分子是参与TGF/Notch-EMT信号通路介导肺癌细胞与骨微环境相互作用的“枢纽”分子。本研究拟通过探讨该分子的表观遗传调控机制及其介导肺癌细胞与骨微环境相互作用的生物学新功能，深入揭示lnc-AF147375及其调控分子在促进肺癌骨转移中的机制。

近年来，多项研究提示长链非编码RNA（lncRNAs）在肿瘤的发生发展过程中起着重要的调控作用，但其在肺癌骨转移中的研究甚少。我们通过lncRNAs表达谱芯片，生物信息学分析，我们筛选到9个对肺癌转移及骨转移起重要作用的关键lncRNA，经过验证后选择了关键分子lncRNA-AF147375。AF147375位于15号染色体上，可能参与了肺癌骨转移相关的TGF-β/NOTCH信号通路。LncRNA-AF147375在癌旁组织中表达高于癌组织，促进了肺癌细胞的侵袭转移；FISH实验提示其在细胞核及细胞质中均有表达，核质分离实验证实其主要表达在细胞核中，进一步机制研究后，我们发现干扰NOTCH3后lncRNA-AF147375的表达上调，而在下游，lncRNA-AF147375可能通过调控TGF-β/c-jun通路参与了肺癌的发生发展，目前进一步的机制实验正在完善中。