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.
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