Lentivirus-mediated reversion-inducing cysteine-rich protein with Kazal motifs gene transfer suppresses pancreatic cancer invasion in vitro.
OBJECTIVE
To investigate the biological functions of reversion-inducing cysteine-rich protein with Kazal motifs (RECK) over-expression in pancreatic cancer cell line Panc-1.


METHODS
This study was carried out in the Department of General Surgery, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China from January to September 2009. The design of the study was to construct a recombinant lentivirus carrying the RECK gene (LV-RECK) to be used to infect Panc-1 cells, and investigate the biological functions of RECK in pancreatic cancer in vitro. The RECK expression was measured by real-time polymerase chain reaction (PCR) and Western blotting. Cell proliferation and apoptosis were examined by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. The ability of invasion was detected using Transwell chambers. The expression and activity of matrix metalloproteinase (MMP)-2 and matrix metalloproteinase-9 (MMP-9) was measured by real-time PCR, Western blotting, and gelatin zymography.


RESULTS
We successfully constructed LV-RECK and proved it can over-express the RECK gene in Panc-1 cells. The RECK over-expression potently suppressed the invasion ability of Panc-1 cells in vitro without affecting cell proliferation and apoptosis. The RECK over-expression potently inhibited the secretion and activity of MMP-2, and the secretion of MMP-9 without affecting the messenger ribonucleic acid, and protein expression of MMP-2 and MMP-9.


CONCLUSIONS
The RECK gene exerts repressive effects on the invasion ability of the pancreatic cancer cell line Panc-1, and it may be a novel therapeutic target for pancreatic cancer.
Tian
Qing-Shui
QS
Department of General Surgery, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China.
Li
De-Chun
DC
D001483:Base Sequence; D015153:Blotting, Western; D003545:Cysteine / Q000737:chemistry*; D017931:DNA Primers; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
D017931:DNA Primers; D011506:Proteins; D003545:Cysteine