MiR-214表达载体的构建及其对肝癌细胞Hep3B增殖的影响

Construction of miR-214 expression vector and its effect on the proliferation of hepatocellular carcinoma cell Hep3B

目的构建pcDNATM6.2-GW/EmGFP-miR-214表达载体,并检测其对肝癌Hep3B细胞增殖的影响。方法以人的基因组DNA为模板,利用PCR扩增miR-214前体序列,构建pcDNATM6.2-GW/EmGFP-miR-214表达载体,序列比对及实时定量PCR验证miR-214表达载体是否构建成功,MTT实验检测miR-214对肝癌Hep3B细胞增殖的影响。结果序列比对及实时定量PCR验证miR-214表达载体构建成功。将成功构建的miR-214表达载体导入肝癌细胞Hep3B,发现其对细胞增殖具有抑制作用。结论本研究成功构建miR-214表达载体并证明miR-214可能参与肝癌发展的过程。

Objective To construct the miR-214 expression vector and explore its effect on the proliferation of hepatocellular carcinoma cell Hep3 B. Methods The precursor of human miR-214 gene was amplified by polymerase chain reaction（ PCR） from the human genomic DNA. The miR-214 expression vector pc DNATM6. 2-GW / Em GFP-miR-214 was constructed and then verified by sequence alignment and real time PCR. Results The pc DNATM6. 2-GW / Em GFP-miR-214 expression vector was successfully constructed and confirmed by the results of sequence alignment and real time PCR. MTT assay showed that miR-214 inhibited the proliferation of Hep3 B cells. Conclusion The miR-214 might be involved in the development of hepatocellular carcinoma.