Alterations in metastatic properties of hepatocellular carcinoma cell following H-ras oncogene transfection

Abstract

Aim: To demonstrate the relationship between H-ras oncogene and hepatocellular carcinoma (HCC) metastasis.

Methods: Activated H-ras oncogene was transfected into SMMC 7721, a cell line derived from human HCC, by calcium phosphate transfection method. Some metastasis-related parameters were detected in vitro, including adhesion assay, migration assay, expression of collagenase IV(c IV ase) and epidermal growth factor receptor (EGFR).

Results: The abilities of H-ras-transfected cell clones in adhesion to laminin (LN) or fibronectin (FN), migration, c IV ase secretion increased markedly, and the expression of EGFR elevated moderately. More importantly, these alterations were consistent positively with the expression of p21, the protein product of H-ras oncogene.

Conclusion: H-ras oncogene could induce the metastatic phenotype of HCC cell in vitro to raise its metastatic potential.

Figure 1

Attachment of different cell clones to LN or FN (^-x ± s_x). A: To increasing concentrations (0, 0.8, 1.6, 2.4 and 3 mg·L^-1) of LN B: To increasing concentrations (0, 5, 10, 20 and 30 mg·L^-1) of FN

Figure 2

Migration ability of representative clones. Subconfluent monolayers of the clones were "wounded" at time 0. The cells were allowed to migration into the cell-free area for 24 h then fixed and stained with crystal violet. A: RC₁; B: RC₂; C: SMMC 7721