Figure 4
Tumorigenicity of ALR reexpressing HCC cells in vivo. (A) Subcutaneously grown tumors in NMRI (nu/nu) mice were analyzed 3 wks after injection of 1 x 106 HepG2-ALR (i-iv) or mock transfected HepG2 cells (v-viii). HE staining of tumors derived from HepG2-ALR cells revealed more epithelial-like cell growth (iii arrow, iv) compared with tumors derived from mock transfected HepG2 cells demonstrating signs of necrotic areas (vii arrow head), cell polymorphism and atypical mitotic figures (viii arrow). Performing immunofluorescence analysis we demonstrated reexpression of EMT markers E-cadherin (i, v) and ZO-1 (ii, vi) in tumors derived from HepG2-ALR cells. (iii, vii, magnification 400x; i, ii, iv-vi, viii, magnification 650x). (B) Western blot analysis confirmed reexpression of EMT markers E-cadherin and ZO-1 as well as reduction of SNAIL expression in tumors originated from HepG2-ALR cells (1 out of 3 experiments).