Abstract
Snail was recently highlighted as a critical transcriptional factor for tumor metastasis. Real time RT/PCR and Western blot analysis demonstrated that Snail mRNA and protein, respectively, were induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in hepatoma cell HepG2. Blockade of gene expression of Snail by antisense oligodeoxynucleotide and/or siRNA technique can prevent not only the TPA-triggered EMT/cell migration and growth inhibition of HepG2 but also TPA-induced down-regulation of E-cadherin and up-regulation of p15(INK4b). Moreover, the TPA-triggered promoter activation of p15(INK4b) was also prevented. On the other hand, two of the HepG2 clone over-expressing Snail, namely S7 and S15, had a scattered fibroblastic morphology and acquired higher motility than parental HepG2. Also, the proportion of G0/G1 phase of S7 and S15 was higher than that of parental HepG2, consistent with the longer doubling time of both cells. Semiquantitative RT/PCR analysis demonstrated a greatly elevated gene expression of Snail accompanied with decreased E-cadherin and increased p15(INK4b) in both Snail-overexpressing cells. On the transcriptional level, p15(INK4b) promoter activity was 2.6-fold higher in S7 as compared with parental HepG2....Continue Reading
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