关键词: 肝细胞癌, 肝细胞核因子4α, 逆转录荧光定量PCR法

Abstract: Objective To establish a quantified reverse transcription-polymerase chain reaction （RT-PCR） for the detection of hepatocyte nuclear factor 4α（HNF4α） mRNA in hepatocellular carcinoma tissues. Method Total RNA was extracted from 16 cancerous tissues of patients with hepatocellular carcinoma（HCC）. The full-length cDNA was obtained by RT-PCR and the products was sequenced after TA cloning. Then,the primer and MGB fluorescent probe were designed for HNF4α mRNA quantification. A one-step RT quantitative PCR assay was established and β-actin mRNA acted as internal control. The HNF4aα mRNA levels in the tissues was calculated. The expression of HNF4α protein was also detected in cancerous tissues by immunohistochemical staining. Results The quantitative detection of HNF4α mRN （V-4α） in HCC tissues was established successfully,and the slope of the standard curve was-3.237 with the correlation coefficient（r） being 0.994. The standard curve of β-actin mRNA was-3.037,and the correlation coefficient（r） was 0.996. The HNF4α mRNA levels were consistent with the HNF4α protein expression in the 16 HCC tissues. Conclusion We successfully establish a quantitative PCR for the detection of HNF4α mRNA in HCC patients,which warrants further study.

Key words: Hepatocellular carcinoma, Hepatocyte nuclear factor 4α, Quantified reverse transcription-polymerase chain reaction