论文部分内容阅读
目的:探索逆转座子LINE-1编码的ORF-1p在肺癌发病过程中的分子机制。方法:利用RNAi技术,在肺癌细胞A549中下调LINE-1编码蛋白ORF-1p,随后对下调后的A549细胞的生物学特征进行细胞增殖(MTT方法)、细胞周期(流式细胞技术)以及集落形成(软琼脂克隆形成试验)等分析,观察细胞生物学特征的改变。并利用报告基因,进一步对细胞周期相关蛋白进行分析。结果:在A549细胞中,下调ORF-1p后细胞的增殖能力明显下降(P<0.05),细胞周期出现S期明显阻滞(P<0.05),肿瘤细胞的集落形成能力明显减弱(P<0.05),p15/p21报告基因表达显示,两种蛋白被显著上调。结论:下调LINE-1基因编码蛋白ORF-1p能够抑制肺癌细胞的生长以及肿瘤的形成。
Objective: To explore the molecular mechanism of ORF-1p encoding retrotransposon LINE-1 in the pathogenesis of lung cancer. METHODS: RNAi was used to down-regulate the ORF-1p protein of LINE-1 in A549 cells. The biological characteristics of A549 cells were then treated by MTT, cell cycle (flow cytometry) Formation (soft agar colony formation assay) and other analysis to observe changes in cell biological characteristics. The reporter gene was used to further analyze cell cycle related proteins. Results: The proliferation of A549 cells was significantly down-regulated after ORF-1p was down-regulated (P <0.05), the cell cycle was significantly blocked in S phase (P <0.05), and the colony forming ability of tumor cells was significantly decreased (P <0.05 ), p15 / p21 reporter gene expression showed that both proteins were significantly up-regulated. Conclusion: Down-regulation of LINE-1 gene encoding ORF-1p can inhibit the growth of lung cancer cells and tumor formation.