Figure 2

Hypoxia induced EMT of HCC cells while reversion occurred under reoxygenation. (A) Morphological changes of hypoxia and reoxygenation-treated SMMC-7721 cells were recorded by light microscope (×200). (B) Immunohistochemical analysis of E-cadherin, N-cadherin and Vimentin expression was performed on both hypoxically and normoxically cultured SMMC-7721 (×200). (C) mRNA levels of E-cadherin, N-cadherin and Vimentin in SMMC-7721 was determined under the conditions of normoxia, hypoxia and reoxygenation by qPCR. GAPDH was used as endogenous reference and mRNA level of SMMC-7721 under normoxia was used as control. Data were shown as mean ± SD of three independent experiments. (D) Protein expression of E-cadherin, N-cadherin and Vimentin in SMMC-7721 cells undergoing normoxia, hypoxia and reoxygenation was determined by immunoblotting. β-actin was used as endogenous control. (E) The numbers of invasive and migrating SMMC-7721 cells undergoing normoxia, hypoxia and reoxygenation was calculated with crystal violet staining. The average numbers of ten random microscopic fields (×400) was recorded in each experiment. Data was shown as mean ± SD of three independent experiments. Representative images of each group were shown.