Effect of mutant p27(kipl) gene on human cholangiocarcinoma cell line, QBC(939).
OBJECTIVE
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.


CONCLUSIONS
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.
Luo
Jian
J
Department of General Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Chen
Yong-Jun
YJ
Wang
Wei-Yu
WY
Zou
Sheng-Quan
SQ
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001650:Bile Duct Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002453:Cell Cycle / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D050760:Cyclin-Dependent Kinase Inhibitor p27; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D014162:Transfection
C436291:CDKN1B protein, human; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D050760:Cyclin-Dependent Kinase Inhibitor p27
Nat Cell Biol. 2001 Mar;3(3):321-4
11231585
Int J Gynecol Pathol. 2004 Jul;23(3):259-64
15213602
BMC Clin Pathol. 2005 Feb 23;5(1):3
15725363
J Urol. 2002 Aug;168(2):766-73
12131366
Int J Gynecol Pathol. 2003 Apr;22(2):168-74
12649672