Figure 5.

The effects of sulfoquinovosyl-acylpropanediol (SQAP) for KYN-2, VHL naturally mutated HCC cell line. (a) Time course of tumor volume changes for subcutaneous KYN-2-bearing mice treated with SQAP (n = 10 per group). All data are represented by mean ± standard deviation (SD). The mice then received the following treatments by intraperitoneal (i.p.) injection: phosphate-buffered saline (control group) or SQAP (20 mg/kg/day, treated group). The treatments were continued for 21 days. (b) Representative Immunohistochemistry images of CD31 staining in KYN-2 are shown. The number of tumor vessels are represented as mean ± SD (n = 10 per group). Scale bar = 50 μm. (c) Representative immunohistochemistry images of TUNEL staining in KYN-2 are shown. The number of apoptotic cells is represented as mean ± SD. (d) Western blots of HIF1α in KYN-2 tumors treated with SQAP. Whole tumors were collected from tumor bearing mice (n = 4 per group). The band densities for each protein were measured and normalized by β-actin. The average band densities are represented as mean ± SD. (e) Western blots of vascular endothelial cell growth factor, HIF1α, HIF2α, VHL, and NFκB for KYN-2 exposed to SQAP under hypoxic conditions are shown. Cells were incubated with SQAP-containing medium for 24 hours, after which the cells were moved to 3% O₂ hypoxic conditions for 24 hours and then lysed with radioimmunoprecipitation buffer. TUNEL, TdT-mediated dUTP nick end labeling.