Aim: To investigate the effects of exogenously mutated p27(kipl) (p27) on proliferation and apoptosis of human cholangiocarcinoma cell line, QBC(939)in vivo.
Methods: Adenviral vectors were used to transfect mutated p27 cDNA into human QBC(939)cell line. Expression of p27 was detected by RT-PCR. Western blot. Cell growth, morphological change, cell cycle, apoptosis and cloning formation were determined by MTT assay and flow cytometry.
Results: The expression of p27 protein and mRNA was increased significantly in QBC(939) cell line transfected with Ad-p27mt. The transfer of Ad-p27mt could significantly inhibit the growth of QBC(939) cells, decrease the cloning formation rate and induce apoptosis. p27 over expression caused cell cycle arrest at G(0)/G(1)phase 72 h after infection with Ad-p27mt.
Conclusion: p27 may cause cell cycle arrest at G(0)/G(1)phase and subsequently lead to apoptosis. Recombinant adenovirus expressing mutant p27 may be potentially useful in gene therapy for cholangiocarcinoma.