抗端粒酶M1RNA核酶真核表达载体构建及肝癌细胞转染的研究

CONSTRUCTION OF EUKARYOTIC VECTOR OF M1RNA RIBOZYME AGAINST TELOMERASE RNA AND TRANSFECTION TO HEPATOCELLULAR CARCINOMA CELL LINES

目的构建抗人端粒酶RNA(hTR)的M1RNA核酶的真核表达载体,并筛选稳定转染核酶的肝癌细胞株。方法设计并应用PCR技术合成hTR模板的M1RNA核酶,应用pEGFPC1载体构建M1RNA核酶的真核表达质粒,应用脂质体LipofectAMINEAM2000转染肝癌细胞系HepG2,应用G418筛选稳定表达核酶的细胞株。结果测序证实hTR的M1RNA核酶基因被正确克隆入真核表达载体pEGFPC1,应用脂质体成功转染肝癌细胞系HepG2。结论成功构建了hTR的M1RNA核酶,并筛选出了稳定表达核酶的肝癌细胞株。

Objective To construct a eukaryotic expression vector of M1RNA ribozyme against telomerase RNA and transfect to hepatocellular carcinoma cell lines. Methods M1RNA ribozyme targeting the RNA template of telomerase was designed. The eukaryotic plasmids of MIRNA ribozyme were constructed using pEGFPCI vector. The plasmids were introduced into hepatocellular carcinoma cells （HepG2） by LipofeetAMINEAM2000. Results The eukaryotic expression vector of M1RNA ribozyme against telomerase RNA was constructed successfully by sequencing analysis. The hepatocellular carcinoma cell lines HepG2 was transfected successfully by M1RNA ribozyme that targeted the RNA component of telomerase. Conclusion The stable transfected cells with M1RNA ribozyme is constructed successfully.