Selection Of High And Low Metastases Cell Lines From B16 Melanoma And Its Molecular Mechanism Of Invasion And Metastases

The course of tumor metastases is very complex. Tumor-bearing host or tumor cells, especially multi-genes, regulate it. To study the mechanisms of tumor metastases and find the anti-tumor targets, we scanned several high/low metastases B16 Melanoma cell lines by the methods of tumor spontaneous lung metastases. The growth or metastases characterizations of these cell line were test in tumor experimental and spontaneous lung metastases model. Then the cell biological methods, such as the test of cell proliferation and 3H-TdR mixing, technology of FCS, and tumor invasion or scatter test, were used to find their differences on growing, cell cycle, responsibility to scatter factor (SF), and invasion. Finally, we also studied their molecular mechanism of high/low metastases using Northern Blotting, RT-PCR, Western Blotting technologies, etc.In these study, we scanned a B16M cell line named H3B16 that had a highest growth speed or most metastases nodules in vivo, an increasing cell proliferation or DNA synthesis and invasion in vitro. It also had a raising proportion of G2-M phase cell and responsibility to Scatter Factor (SF). We detected and found a higher expression of C-met, Ets and VEGFR gene in high metastases H3B16 cells than lowmetastases B16M cells in mRNA or protein level. The phosphorylation of ERK and Akt, which are key step in down-stream of HGF/C-met signal transduction pathway, were increased in H3B16 cell line. The activities of MMP2/9 in H3B16 were promoted. We also found a high expression of IL-18, Tec, or Annexinl and decreasing expression of nm23 in H3bl6M cell line.It seems suggested that changed expression of c-met or Ets, or VEGFR and nm23 leaded a exceed proliferation and high invasion or metastases in H3b16M cells through activating of its down-stream signal pathway. The role of changed IL-18 or Tec gene expression in H3B16M will be studied lately.