Figure 2. Characterization and Evaluation of anti-tumor effects of a SMI of β-hydroxylase (MO-I-1151) on the HCC phenotype.

(A) MTT assays showed relative absorbance at each concentration (ranging from 1.25 to 10 µM) of MO-I-1151, MO-I-1100 and other candidate β-hydroxylase inhibitors (MO-I-1171, 1178, 1351, 2751, 5007, 5008, 5009, 5010, 5018 and 5019) which had no biologic activity. The assay was performed after 24-hour’s exposure to DMSO or SMIs in FOCUS HCC cells. [Blue line was MO-I-1000 (the first generation SMI). red line was MO-I-1151 (the second generation SMI). Black lines represented other compounds that had no effect.] (B) MTT assays showed relative absorbance at each time point exposed to DMSO or different concentrations of MO-I-1151 (from 1.25 to 10 µM) for 5 days. (C) The bar graphs indicated relative colony numbers after 3-week exposure to DMSO or MO-I-1151 (2.5 or 5 µM). Representative photographs of anchorage-independent colony formation were shown. (D) Representative photographs of SA-β-gal staining after 24-hour’s exposure to DMSO or MO-I-1151 (5 µM) (left panel, 200×). The SA-β-gal activity was quantified and expressed as percent of SA-β-gal-positive cells (right panel). Mean ± S.D. of triplicate independent experiments. *P < 0.05, **P < 0.01 compared to DMSO treated cells.