LncRNA-AK123073对肺癌细胞改造骨微环境的作用和机制研究

Lung cancer is the most common malignant tumor in terms of both incidence and mortality worldwide, and distant metastasis is considered as the leading cause of lung cancer. 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 of patients and bring trouble to clinical treatment. So, it is necessary to clarify the mechanism of bone metastasis of lung cancer and develop new more effective treatment against bone metastasis. Our previous works showed various genes 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 path ways. Our previous works firstly indicated lncRNAs are pivotal factors in the remodeling of lung cancer cells to bone microenvironment, but detailed mechanism is not clear. In this study, series of molecular biological approaches will be employed to further investigate the role of lncRNA candidates lncRNA-AK123073 with related regulational moleculars 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.

肺癌是发病率和死亡率高的恶性肿瘤，约1/3肺癌患者会发生骨转移而引起剧烈疼痛、骨折和截瘫等，严重影响患者的生存质量，是临床亟需解决的难题。对肺癌骨转移机制的研究是探讨新的治疗方法的前提。申请者长期致力于肺癌骨转移机制研究，前期研究中发现肺癌细胞对骨微环境的改造在骨转移发生发展中发挥着至关重要的作用。LncRNA具有调控癌细胞信号网络的功能，我们前期从临床骨转移肺癌组织和动物模型，筛选获得了9个明显差异表达的lncRNAs，并首次研究发现lncRNA-AK123073与肺癌骨转移有关，其可能是肺癌细胞改造骨微环境调控网络中的一个“枢纽”分子。本研究拟基于前期基础，通过下游靶基因筛选、表观遗传调控、骨微环境改造功能研究等，对lncRNA-AK123073及其调控分子在肺癌细胞改造骨微环境中的作用和机制进行深入研究，为后续研发肺癌骨转移新药物奠定基础。

肺癌是发病率和死亡率最高的恶性肿瘤，骨转移是肺癌患者最常见的远处转移类型，骨相关事件严重影响着肺癌患者的生存质量和预后，对肺癌骨转移机理的研究是探讨新的肺癌骨转移治疗方法的重要前提。本研究组前期采用高通量lncRNA microarray技术，针对临床肺癌标本库和特异性肺癌骨转移细胞株，进行了lncRNA芯片筛选，获得了9个肺癌骨转移组织细胞中差异表达的lncRNAs分子，并对其中lncRNA-AK123073进行了初步研究，发现lncRNA-AK123073可通过调控转移相关分子参与了肺癌骨转移的发生，干预lncRNA-AK123073的表达有抑制体内骨转移发生的作用。进一步的研究发现其可能通过影响骨微环境而促进了肺癌骨转移的发生。在本项目中，我们运用高通量lncRNA芯片发现LncRNA-AK123073在发生骨转移的肺癌组织中表达量显著降低，进而我们设计过表达LncRNA-AK123073的慢病毒载体，感染A549和H1299细胞，成功构建了过表达LncRNA-AK123073的细胞，并通过CCK8细胞增殖实验、克隆形成实验、损伤刮擦实验和Transwell侵袭实验证实，过表达LncRNA-AK123073能抑制非小细胞肺癌细胞A549和H1299细胞的增殖、克隆形成、迁移和侵袭能力；进一步通过流式细胞技术研究LncRNA-AK123073对非小细胞肺癌细胞凋亡和周期的影响，结果显示过表达lncRNA-AK123073能促进A549和H1299细胞的凋亡发生比率，G1期的细胞数量减少，而G2/M期的细胞比率相对增加。通过利用PhyloCSF的方法，提示为AK123073非编码RNA。通过核质分离实验，发现lncRNA-AK123073主要位于胞核，为进一步判断该lncRNA的机制提供了依据。动物体内研究和分子调控机制拟在2020年度进一步完善。