A novel human DNA sequence with tumor metastasis suppressive activity

Abstract

To isolate human tumor metastasis suppressive DNA sequence and to study the molecular mechanisms regulating tumor metastasis.A mouse lung adenocarcinoma cell clone 12 derived from its parent cell line LM2, which had been transduced with normal human genomic DNA, was previously reported. Compared with LM2, the metastatic potential of clone 12 was very much decreased. Clone 12 was used in this study to amplify the human DNA fragments by Inter Alu PCR technique. The human DNA fragments obtained were then transfected into LM2 cells and their malignant phenotype was observed in vitro and in vivo, and compared with that of the untransfected LM2 cells.Three human DNA fragments of 700, 500 and 300 bp were isolated. DNA sequencing revealed that the 700 bp fragment does not show homology with hitherto reported genes and was accepted by the GenBank (pt 712 U67835). In vitro proliferation and colony formation in soft agar of the 700 bp fragment-transfected LM2 cells were significantly inhibited as compared to the untransfected LM2 cells. Upon subcutaneous inoculation to syngeneic T739 mice, the 700 bp-transfected LM2 cells grew more slowly and smaller tumors developed compared to the un-transfected ones. Moreover, lung metastasis was not found in 6 of 10 mice inoculated with the 700 bp-transfected LM2 cells, while it was found in 9 of 10 mice inoculated with the un-transfected LM2 cells. The difference was statistically significant (P < 0.001). The frequency of lymph node metastasis was also statistically different between the 2 groups of mice.The newly isolated 700 bp human DNA fragment may be a metastasis suppressor gene of malignant tumor.