Decreasing Pin1 suppressed the properties of migratory and invasive in colorectal carcinoma SW620 cells

The Chinese-german Journal of Clinical Oncology(2010)

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摘要
Objective The aim of our study was to investigate the effect of Pin1 on the expression of MMP-2 and MMP-9 in human colorectal carcinoma SW620 cells. Methods We constructed a eukaryotic expression vector of RNA interfering (shRNA) for Pin1 gene (pGenesil-1-Pin1), and then observed its expression in SW620 cells by Western blotting. The cells motility were tested by wound healing assay and Boyden chamber assay. The protein levels and activity of MMP-2 and MMP-9 were tested by Western blotting and Gelatin zymography in SW620 cells after transfected with pGenesil-1-PIN1. Results pGenesil-1-PIN1 was successfully constructed, which was confirmed by sequencing. Silencing the Pin1 by RNAi significantly decreased the cells motility from 96.4 ± 3.9 per field (× 10 objective) to 52.7 ± 4.4 per field ( P < 0.05, Student’s t -test) for SW620 cells transfected with pGenesil-1-PIN1 (SW620/p-shRNA) in Boyden chamber assay, and reduced the MMP-2 and MMP-9 expressions and activity in SW620 cells. The protein relative levels of MMP-2 were 0.32 ± 0.04 for SW620/p-shRNA, and 0.76 ± 0.03 for SW620/p-Con; MMP-9 were 0.41 ± 0.09 for SW620/p-shRNA, and 0.94 ± 0.07 for SW620/p-Con ( P < 0.05). Conclusion Inhibited Pin1 expression may contribute to the suppression of the invasive and metastatic capacity of colon cancer cells in vitro .
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关键词
rnai,pin1,mmps,metastasis and invasion,colorectal cancer,cell motility,colorectal carcinoma,colon cancer
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