engleski

Tumor metastasis supressor gene nm23-H1 promotes transcription of matrix metalloproteinase-13 in CAL 27 cells

The human nm23-H1 has been discovered as a tumor metastasis suppressor on the basis of its reduced expression in melanoma cell lines with low vs. high metastatic potential. It encodes for one of the two subunits of the nucleoside-diphosphate kinase. Besides the apparent role in the maintenance of the cells NTP pool it has become obvious that nm23 plays a key role in different cell processes: proliferation, differentiation, cell signaling etc., and has some other biochemical properties such as DN-ase or kinase activity. The involvement of nm23-H1 in these processes must still be elucidated. Since obviously dealing with a multifunctional enzyme our goal was to take advantage of the recently developed microarray technology. We constructed CAL 27 cell clones (SCC of the tongue) constitutively expressing EGFP-nm23-H1. The microarray analysis was preformed using Affymetrx U133A expression chip according to the manufacturer's instructions. The results revealed 138 genes had elevated while 103 hadlowered expression compared to the control. The genes with changed expression patterns could roughly be clustered in TGFb and matrix metalloproteinase pathway, phosphatidil-inositole signaling pathway the proteasome machinery or the cell-cycle. Our attention was particularly driven to the genes of the TGFb pathway and the matrix metalloproteinase 13 (MMP13) whose expression was significantly elevated in GFP-nm23-H1 expressing cells. The results were confirmed by real-time PCR and Western blot analysis. Since the MMP family is actively involved in the degradation of the extracellular matrix, cell migration and metastasis formation, our future experiments will be focused on the interaction of nm23-H1 and the MMP13.

nm23-H1; MMP13; cDNA microarray; CAL 27 cells

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