miR-1调控MALAT-1表达在化学致癌物乌拉坦诱发肺细胞癌变中作用

Long non-coding RNA(LncRNA) is a recently discovered kind of endogenous nucleic acid molecules, which play a role in negative regulation of gene expression . The expression of lncRNA is regulated by microRNA(miRNA). This study will explore the mechanism of lung cancer induced by environment chemicals from new perspective based on miRNA and lncRNA interaction. This project used malignant transformation of human bronchial epithelial cells induced by urethane as model, established the function research system of miR-1 and selected molecular events closely related to the development of lung cancer: proliferation, apoptosis, cell cycle, invasion and migration, to study the role of miR-1 during malignant transformation of pulmonary epithelial cells induced by urethane. This project will confirm the relationship between miR-1 and lncRNA MALAT-1 by using luciferase gene reporter system. Utilizing means of siRNA or over-expression and analyze the effect of inhibiting MALAT-1 expression or up regulation of MALAT-1 expression on the biological behavior of transformation cells, to analyse effect of MALAT-1 on the biological behavior of transformed cells and explore the functional significance of pulmonary carcinogenesis induced by urethane in the progress of occurence and development. The results of this study not only provide a new idea for research on carcinogenic mechanism of environmental chemicals，meaningfully, provide experimental data for intervention and prevention of lung cancer in the early stage.

长链非编码RNA(lncRNA)是近年发现的细胞内对基因表达具有重要调控作用的内源性核酸分子, lncRNA的表达受microRNA(miRNA)的调控。本研究从miRNA与lncRNA相互作用的新视角探讨环境化学物诱发的肺癌机制。本项目以致癌物乌拉坦诱发人永生化支气管上皮细胞恶性转化，建立miR-1的功能研究体系，选择同肺癌发生发展密切相关的分子事件：增殖、凋亡、细胞周期、侵袭、迁移，研究miR-1在乌拉坦诱发肺上皮细胞恶性转化过程中的作用。通过荧光素酶基因报告系统，确认miR-1与长链非编码RNA MALAT-1靶向关系。运用干扰抑制或过表达等手段，分析下调或上调MALAT-1对转化细胞生物学行为的影响，探索miR-1调控MALAT-1表达在乌拉坦诱发肺癌发生与发展过程中的功能意义。本研究不仅将为环境化学物致癌机制研究提供新思路，更有意义的是为肺癌的早期基因干预防治提供实验研究数据。

本研究以典型环境化学致癌物为受试物，处理永生化人支气管上皮细胞，建立环境化学物诱导人支气管上皮细胞恶性转化模型，探索长链非编码RNA(lncRNA) MALAT1及 microRNA-1在环境化学物致癌过程中的作用及分子机制。运用实时定量PCR法检测了致癌物急或慢性处理的上皮细胞（HBE、BEAS-2B及A549）中MALAT1及microRNA-1等非编码RNA分子的表达. 通过脂质体转染及siRNA转染技术，分析了细胞中长链非编码RNAMALAT1及microRNA的表达调控关系。发现lncRNA MALAT1等lncRNA可调控细胞增殖等恶性表型。在致癌物苯并芘短期处理上皮细胞之后，诱导了MALAT1表达改变、促炎因子表达升高及迁移与侵袭能力增强，研究表明MALAT1可能通过调控炎症因子表达牵涉苯并芘致癌。本项目研究lncRNA MALAT1等长链非编码RNA分子在环境致癌物诱发肺癌中的作用，为肺癌的早期基因干预防治提供了实验研究数据。