Abstract: Objective To develop a real-time fluorescent quantitative polymerase chain reaction（RT-PCR）for the detection of cortactin gene expression in human hepatocellular carcer（HCC）tissue. Methods Total RNA was extracted from human liver cancer tissue samples.Cortactin and GAPDH gene were amplified by RT-PCR and the products of each gene were inserted in a pMD18-T vector to plot standard curves by real-time PCR with SYBR Green I.Five human liver cancer tissue samples were detected based on the standard curves and the threshold cycle（Ct）value was analyzed to evaluate the precision,linearity and reproducibility of this method. Results pMD18-T（+）/CTTN and pMD18-T（+）/GAPDH plasmids were successfully constructed.When the concentration of temples was 1.6×10³-1.6×10⁷ and 1.6×10⁴-1.6×10⁷ copies/μL,prespectively,cortactin and GAPDH standard curves had a good linearity（R²=0.996 and 0.985）.The repeatability of two genes was 0.11%-2.25% and 0.75%-3.63%（coefficient of variability）respectively within run and 0.83%-1.48% and 0.47%-1.36%（coefficient of variability）respectively between days.No statistical difference was observed between days（P＞0.05）. Conclusion The RT-PCR developed in this study lays a methodological foundation for the study of CTTN gene expression level in human HCC tissue.