LINE-1ORF-1p过表达对肝癌细胞株SMMC7721增殖和锚定非依赖性生长的影响

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;lt 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.