Effects of recombinant human canstatin protein in the treatment of pancreatic cancer

Figure 1 Amplification products of human canstatin. Lane M: DNA marker; Lane 1: RT-PCR products.

Figure 2 Restriction analysis of plasmid pUCm-T/canstatin. Lane M: DNA marker; Lane 1: plasmid pUCm-T/ canstatin; Lane 2: pUCm-T/ canstatin digested by BamHI; Lane 3: pUCm-T/canstatin digested by HindIII.

Figure 3 Restriction analysis of plasmid pET-22b(+)/canstatin. Lane M: DNA marker; Lanes 1 to 7: plasmid DNAs of seven selected colonies digested by both BamHI and HindIII.

Figure 4 SDS-PAGE gel electrophoresis of purified protein. Lane M: protein marker; lane 1: total bacterial protein; lane 2: 10 mmol/L imidazole elution; lane 3: 25 mmol/L imidazole elution; lane 4: 50 mmol/L imidazole elution; lane 5: 125 mmol/L imidazole elution; lane 6: 250 mmol/L imidazole elution.

Figure 5 In vivo antitumor effects in a xenograft model. The in vivo antitumor effect of the canstatin was analyzed in a SW1990 human pancreatic cancer cell orthotopic xenograft model. A: Tumor volumes in different groups: Canstatin 5 mg/kg or 10 mg/kg treatment showed a significantly stronger anti-tumor effect, compared with the PBS treated group (^bP < 0.01); B: MVD in xenograft models: significant inhibition of angiogenesis was observed in groups treated with canstatin 5 mg/kg or 10 mg/kg compared with that in the control group (^aP < 0.05, ^bP < 0.01, respectively); C: CD34 immunohistochemistry in group treated with PBS (× 200): tumor sections showed extensive angiogenesis; D: CD34 immunohistochemistry in group treated with canstatin (× 200): tumor sections showed obviously decreased new vessels with a small focus of necrosis.