摘要
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 investigate the mechanism of LINE-1 encoded ORF-1p in the tumorigenesis of lung cancer.Methods:The down-regulation of ORF-1p was conducted by RNAi approach and the features of transfected A549 cells were characterized by the approaches of MTT,flow cytometry and soft-agar anchored colony formation.Results:With the constructs of recombinant plasmids,LINE-1 encoded ORF-1p was down-regulated significantly in transfected A549 cells.The characters of transfected A549 cells were appeared to change as follows:Cell proliferation was significantly reduced,which was probably attributed to up-regulation of p21 and p15;cell cycle exhibited S phase arrest(P0.05);colony formation was significantly attenuated.Conclusion:The down-regulation of ORF-1p can attenuated the A549 characters significantly upon its tumor formation ability.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第6期14-20,共7页
China Biotechnology