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. 2019 Sep 19;9(23):6745–6763. doi: 10.7150/thno.34674

Figure 5.

Figure 5

CA terminated Huh7 and H446 tumor growth in vivo. (A) Tumor growth inhibition by CA (ip, 25 or 50 or 200 mg/kg/d) in NOD/SCID mice inoculated with Huh7 (left) or H446 (right) cells. Arrows indicate the xenograft tumor entered non-growth phase, after treatment with CA at different doses. Tumor tissues taken from the CA-treated mice largely lost tumorigenicity after re-implantation of the tumor cells into naïve mice for 28 days (inserts, n = 4). Data are presented as mean ± SD. * p < 0.05 and *** p < 0.001 (CA treated versus saline control). (B) Tumor weight in the NOD/SCID mice inoculated with Huh7 (left) or H446 (right) cells followed by CA treatment (ip, 25 or 50 or 200 mg/kg/d) and saline treatment used as a control. CA treatment (ip, 25 mg/kg/d or 50 mg/kg/d or 200 mg/kg/d) yielded 35.91%, 76.34% or 77.33% tumor weight inhibition of Huh7 tumors and 36.90%, 74.14% or 74.72% tumor weight reduction of H446 tumors, respectively. (C) mRNA expression of the study genes in Huh7 (upper) or H446 (lower) tumor tissues taken from the mice treated with CA (25 mg/kg/d) was examined by qRT-PCR (n = 6). (D) Protein expression of c-Myc and p21 in Huh7 (left) or H446 (right) tumors was analyzed by Western blotting (n = 6, upper panel). Density scanning results are presented as well (lower panel). (E) Expression levels of miR-17 family members in Huh7 (upper) or H446 (lower) tumors taken from the CA-treated mice was assessed by qRT-PCR (n = 6). (F) The nucleus sumoylation of c-Myc in Huh7 or H446 tumors was measured using EpiQuikTM Protein Sumoylation Assay Kit (n = 6). (G) Body weight measurement on day-7 after CA injection (ip; 1000, or 500, or 250 mg/kg/d; n = 10 for each group). (H) Liver and kidney function remained normal after CA treatment. AST: aspartate transaminase (U/L); ALT: alanine aminotransferase (U/L); BUN: blood urea nitrogen (mM); CRE: creatine (μM). The data are presented as mean ± SEM. * p < 0.05, ** p < 0.01 and *** p < 0.001 (CA treated versus normal saline control). (I) Cmax of CA in rats was measured after ip injection of CA (75 mg/kg). Three rats were used for each time point. Cmax, the maximum concentration in blood. Data are presented as mean ± SD.