Expression pattern of SFRP1 in HCC specimens by RT-PCR and immunohistochemical staining. (A) Representative results of semi-quantitative RT-PCR of SFRP1 from 24 pairs of HCCs (C) and corresponding adjacent non-cancerous livers (N), where β-actin was employed as an internal control. Each PCR was generally performed in 32 thermal cycles and PCR products were visualized after electrophoresis through 2% agarose. The length of PCR product of SFRP1 and β-actin are 328 bp and 295 bp, respectively. (B) To confirm the absence of influences of genomic DNA contamination, 6 Paired HCCs and non-HCCs were selected randomly to detect the SFRP1 and beta-actin expression by RT-PCR. Experiments were performed by using RT (RT+) or no RT (RT-) in each sample. (C) Real time RT-PCR analysis of SFRP1 was carried out on 46 paired HCCs and adjacent non-cancerous livers. For each sample, the relative mRNA level of SFRP1 was normalized based on that of β-actin. The line within each box represents the median -ΔCt value; the upper and lower edges of each box represent the 75th and 25th percentile, respectively; the upper and lower bars indicate the highest and lowest values determined, respectively. * indicates p value <0.001. (D) Representative immunohistochemical staining of a pair of HCC specimen and corresponding non-cancerous liver with anti-SFRP1 antibody. The nuclei were countered stained with hematoxylin. (E) Expression pattern of SFRP1 was evaluated in HCC cell lines, fetal and adult normal livers through RT-PCR, where β-actin was used as a loading control.