β-catenin accumulation in nuclei of hepatocellular carcinoma cells up-regulates glutathione-s-transferase M3 mRNA

Figure 1 Identification of differentially displayed cDNA fragments from paired hepatocellular carcinoma cells (T) and nontumorous liver tissues (N). A: Differentially displayed PCR products (“I” and “II”) amplified with AP-10 primer and T₁₂MA. Lanes 1-3 denote the three B6C3 F1 mouse samples, respectively; B: Recovered “I” and “II” from the dried DNA sequencing gel reamplified by PCR. MW lane shows the pUC118 DNA fragments cut by HapII (a restriction enzyme) as molecular weight markers; C: Nucleotide sequences of the bands shown in A. Flanking sequences of T₁₂MA primers are underlined. The two insert-containing fragments were sequenced and identified as gene fragments of GLNS and GSTM3 in comparison with those of nucleotide in GenBank.

Figure 2 Levels of glutamine synthetase and glutathione-s-transferase M3 mRNA and expression of β-catenin. A: T denotes surgically resected B6C3F1 mouse hepatocellular carcinoma (HCC) tissue samples and N denotes matched nontumorous liver tissue samples; B: Representative immunofluorescence photomicrographs for β-catenin (red photomicrographs) in HCC tissue samples. B1 denotes negative β-catenin staining in nuclei of HCC cells, B2 denotes positive β-catenin staining in nuclei of HCC cells, and white arrows indicate β-catenin detected in nuclei of HCC cells. Bars: 50 μm. GLNS: Glutamine synthetase; GSTM3: Glutathione-s-transferase M3.

Figure 3 Identification of β-catenin/Tcf-Lef consensus binding sites with three β-catenin/Tcf-Lef consensus binding site sequences [5'-(A/T)(A/T) CAAAG-3'] located at the nearby upstream domain of mouse GSTM3 by searching GenBank.

Figure 4 Total glutathione-s-transferase activity (A) and glutathione-s-transferase M3 mRNA expression (B) in HepG2 cells. ^aP < 0.05, ^bP < 0.01 vs TWS119 at 24 h by one-way analysis of variance followed by Bonferroni’s test.