摘要
目的研究逆转座子L1编码蛋白ORF-1p的表达对肝癌细胞株SMMC7721细胞增殖的影响。方法利用带Flag标签的L1 ORF1表达载体,转染SMMC7721细胞;采用Western blot法检测反转座子L1 ORF1编码蛋白ORF-1p的表达;采用软琼脂成集落实验和相关报告基因检测ORF-1p表达对肝癌细胞p53、p15和p21活性的影响。结果在SMMC7721细胞中,Flag-ORF-1p表达能够显著促进该细胞的增殖(P<0.05),增强该细胞的锚定非依赖性生长(P<0.05),降低p53、p15和p21报告基因的活性。结论 L1基因编码蛋白ORF-1p能够促进肝癌细胞的生长、浸润以及肿瘤的形成。
Objective To investigate the effects of L1 encoded ORF-1p on the proliferation of SMMC7721 cells.Methods Recombinant expression vector pcDNA3.0-Flag-ORF1 was used to transfect SMMC7721 cells;Expression of ORF1p in the transfected cells was detected by Western blotting;Proliferation and anchor-independent growth of the cells transfected with Flag-ORF1p vector were detected by MTT assays and soft-agar anchored colony formation assays;Transcriptional activity of p15,p21 and p53 reporter genes was analyzed by luciferase reporter system.Results Flag-ORF-1p could enhance the proliferation and anchor-independent growth ability of SMMC7721 cells;It also reduced the transcriptional activity of p15,p21 and p53 luciferase report genes.Conclusion ORF-1p of L1 might enhance the SMMC7721 cell proliferation and anchor-independent growth through regulation of some genes.
出处
《实用肝脏病杂志》
CAS
2011年第5期323-326,共4页
Journal of Practical Hepatology
基金
国家科技重大专项课题(2008ZX100022018)
首都医学发展基金课题(2009-2041)