论文部分内容阅读
目的研究反转座子LINE-1编码蛋白LINE-1 ORF-1p对肝脏肿瘤细胞系以及肝源永生化细胞系增殖能力的影响。方法构建针对LINE-1 ORF-1编码序列的siRNA表达载体(LINE-1 ORF-1p psilence2.1-U6-siRNA),转染肝脏肿瘤细胞系Bel-7402、SMMC-7721、HepG2和肝源永生化细胞系LO2。采用Western blotting检测LINE-1 ORF-1p siRNA表达载体对LINE-1 ORF-1编码蛋白LINE-1 ORF-1p表达的影响,采用MTT法检测LINE-1 ORF-1p表达受抑对细胞增殖的影响。结果在肝脏肿瘤细胞HepG2、Bel-7402、SMMC-7721和肝源永生化细胞LO2中,转染LINE-1 ORF-1p siRNA表达载体均能降低LINE-1 ORF-1p的表达水平,并从第3天起显著抑制前述细胞的增殖(P<0.05),其抑制率分别为63%、51%、40%、43%。结论抑制LINE-1 ORF-1p蛋白表达后可使肝脏肿瘤细胞系和永生化细胞系的增殖受抑。
Objective To investigate the effect of LINE-1-encoded protein LINE-1 ORF-1p on the proliferation of hepatic tumor cell lines and immortalized hepatocytes. Methods The siRNA expression vector (LINE-1 ORF-1p psilence2.1-U6-siRNA) targeting to ORF-1 of LINE-1 gene was constructed and transfected into liver tumor cell lines Bel-7402, SMMC-7721, HepG2 and immortalized liver Cell line LO2. The effect of LINE-1 ORF-1p siRNA expression vector on the expression of ORF-1 and ORF-1 in LINE-1 ORF-1 protein was detected by Western blotting. The effect of ORF-1p expression on the proliferation was detected by MTT assay . Results The expression of LINE-1 ORF-1p in the liver tumor cells HepG2, Bel-7402, SMMC-7721 and liver-derived immortalized L02 cells transfected with the vector of LINE-1 ORF- 3 days significantly inhibited the proliferation of the aforementioned cells (P <0.05), the inhibition rates were 63%, 51%, 40%, 43%. Conclusion Inhibition of LINE-1 ORF-1p protein expression can inhibit the proliferation of liver tumor cell lines and immortalized cell lines.