Effects of HepaCAM gene transfection on invasiveness of renal cancer cells
10.3781/j.issn.1000-7431.2008.12.003
- Author:
Chun-Hua XUN
1
Author Information
1. Department of Laboratory
- Publication Type:Journal Article
- Keywords:
Cell adhesion molecules;
Genes, hepaCAM;
Kidney neoplasms;
Neoplasm invasion;
Transfection;
Tumor cells, cultured
- From:
Tumor
2008;28(12):1029-1033
- CountryChina
- Language:Chinese
-
Abstract:
Objective: This study was to evaluate the effects of hepaCAM (hepatocyte cell adhesion molecule) on invasiveness of human renal cell carcinoma (RCC) cell line 786-0 and explore the underlying mechanism. Methods: A recombinant plasmid containing hepaCAM cDNA (hepaCAM-pEGFPN2) was transfected into 786-0 cells, then the positive cell clones were selected with G418 and amplified. The 786-0 cells transfected with pEGFPN2 and untreated 786-0 cells were as controls. The mRNA expressions of hepaCAM and matrix metalloproteinase (MMP)-2 and MMP-9 before and after stable transfection are detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time fluorescence PCR. The activities of MMP-2 and MMP-9 were analyzed by gelatin zymography. The invasiveness of 786-0 cells was determined by Transwell migration assay in vitro. Results: The mRNA transcription of hepaCAM was significantly increased in 786-0 cells after stable transfection of hepaCAM (P <0.01). The mRNA expressions and activities of MMP-2 and MMP-9 in 786-0 cells were markedly inhibited by hepaCAM (P <0.05). The transmembrane migration capacity of cells was significantly reduced after stable transfection of hepaCAM (P <0.01). Conclusion: HepaCAM inhibits the activities of gelatinase by down-regulating the expressions of MMP-2 and MMP-9, thereby preventing the invasiveness of renal cell carcinoma 786-0 cells.
