[Adenovirus-mediated overexpression of KAI1 suppresses sphingosine kinase activation and metastasis of pancreatic carcinoma cells].
OBJECTIVE
To investigate influence of KAI1 expression on the pancreatic carcinoma cell migration and its mechanisms.


METHODS
We construct an adenoviral vector carrying the full length cDNA of KAI1 (Ad-KAI1). The Ad-KAI1 virus was packed and amplified in 293 cells and was purified by Adeno-X Virus Purification Kit. The adenovirus-mediated KAI1 expression in human pancreatic carcinoma cell line (PANC I) was detected by flow cytometric analysis. Sphingosine kinase (SPK) activation was determined by [gamma-(32)P] incorporation. And cell migration was assayed by using transwell.


RESULTS
We constructed Ad-KAI1 vector successfully. The 86.3% of pancreatic cancer cells PANC I were identified and infected by Ad-KAI1 constructed by us. At same time, we found out higher concentration of SPK in the PANC I cancer cells without infection of Ad-KAI1 using RT-PCR. Overexpression of KAI1 significantly inhibited the activity of SPK and the migration ability of the PANC I cells, which was induced by activity of SPK.


CONCLUSIONS
Adenovirus-mediated overexpression of KAI1 suppresses sphingosine kinase activation and migration of pancreatic carcinoma cells. This research is helpful for elucidating the anti-metastasis mechanisms KAI1 in pancreatic carcinoma cells.
Guo
Xiao-zhong
XZ
Department of Gastroenterology, Shenyang General Hospital of PLA, Shenyang 110016, China. gxzsw@pub.sy.ln.cn
Zhang
Wei-wei
WW
Wang
Li-sheng
LS
Lu
Zhuo-zhuang
ZZ
Wang
Hua
H
Xu
Jian-hua
JH
Tian
Hong
H
D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D006801:Humans; D051861:Kangai-1 Protein / Q000235:genetics*; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D014161:Transduction, Genetic; D014162:Transfection; D000099157:Sphingosine Kinase
D051861:Kangai-1 Protein; D017853:Phosphotransferases (Alcohol Group Acceptor); D000099157:Sphingosine Kinase