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a                        b
           a                       b














                                                               c                        d

           c                       d














                                                               e                        f
                                                              Figure 3: Photomicrographs of image analysis of immunohistochemically stained
                                                              EAC-sections for Bcl-2 (b), p53 (d) and TdT (f) of EAC-bearing mice treated with
           e                       f                          DADS as compared to their respective EAC-bearing mice controls (a, c and e).
          Figure 2: Photomicrographs of immunohistochemically stained EAC-sections.   The black colored areas refl ect the yellowish brown stained areas. The fi gures
          Images respectively show the greater amount of Bcl-2 (arrow; yellowish brown   depict a decreased expression of Bcl-2 and an increased expression of p53 and
          color) in the cytoplasm of EAC-cells in control mice (a, ×100) as compared with   TdT as a result of treatment with DADS. EAC: Ehrlich ascites carcinoma; DADS:
          mice treated with DADS (b, ×100), the higher amount of p53 (arrow; yellowish   diallyl disulfi de; TdT: terminal deoxynucleotidyl transferase
          brown color) in the cytoplasm and nuclei of EAC-cells in mice treated with
          DADS (d, ×100) than in EAC-bearing control mice (c, ×100), and the higher level   potentially elevated in EAC-bearing mice treated with DADS
          of TdT (arrow; yellowish brown color) in the nuclei of EAC-cells in mice treated with
          DADS (f, ×100) than in EAC-bearing control mice (e, ×100). EAC: Ehrlich ascites   as compared with those of EAC-bearing control mice.
          carcinoma; DADS: diallyl disulfi de; TdT: terminal deoxynucleotidyl transferase
                                                              Effects on EAC cells in vitro
          As indicated in Figure 4, the area in pixels, percent area   Incubation of EAC-cells (2.5 × 10 /mL suspended in PBS)
                                                                                           5
          and intensity of yellowish brown color of immunoreactive   with 0, 6.25, 12.5, 25, 50, and 100 μg/mL of DADS (dissolved
          anti-apoptotic marker, Bcl-2, were remarkably decreased
          in EAC-bearing mice treated with DADS as compared to   in DMSO) for 2 h produced 0, 16.5%, 35%, 70%, 90%,
          those of EAC-bearing control mice. In contrast, the area in   and 100% inhibition of cell viability, respectively. Thus,
          pixels, percent area and intensity of yellowish brown color   the effect seemed to be dose dependent and IC50 was
          of immunoreactive apoptotic markers, p53 and TdT were   19.500 μg/mL [Table 3].



               Hepatoma Research | Volume 1 | Issue 2 | July 15, 2015                                        71
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