Premium
Targeting the HGF/SF receptor c‐met using a hammerhead ribozyme transgene reduces in vitro invasion and migration in prostate cancer cells
Author(s) -
Davies Gaynor,
Watkins Gareth,
Mason Malcolm D.,
Jiang Wen G.
Publication year - 2004
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.20068
Subject(s) - c met , hammerhead ribozyme , hepatocyte growth factor , biology , receptor tyrosine kinase , transgene , microbiology and biotechnology , ribozyme , transfection , prostate cancer , in vitro , cancer research , receptor , medicine , cancer , gene , rna , biochemistry , genetics
Background Hepatocyte growth factor scatter factor (HGF/SF) elicits a number of biological activities including invasion and migration through activation of its tyrosine kinase receptor c‐Met. Over expression of c‐Met has been implicated in prostate cancer development and progression. This study examined the effect of a ribozyme transgene, designed to inhibit human c‐Met expression, and its impact on in vitro invasion and migration in prostate cancer. Methods A transgene (Met 560) consisting of U1 snRNA, hammerhead ribozyme, and antisense was cloned into a modified pZeoU1‐EcoSpe vector and transfected into DU‐145 cells. The effect of HGF/SF was tested on prostate cancer cells whose expression of c‐Met had been blocked by way of a ribozyme transgene. Results Met 560 stable transfectants (DU‐145 +/+ ) manifested a complete loss of c‐Met expression at mRNA and protein levels. In contrast, control plasmid (DU‐145 +/− ) and wild‐type DU‐145 cells (DU‐145 −/− ) had similar levels of c‐Met expression. HGF/SF significantly increased the in vitro invasiveness (mean 47.71 ± SE 7.75; P < 0.01 vs. control 24.14 ± 1.34), and migration (mean 48.44 ± SE 3.51; P < 0.01 vs. control 22.95 ± 1.47) of DU‐145 −/− cells, respectively. Similarly, HGF/SF also increased the invasion (62.33 ± 6.34; P < 0.001 vs. control 24.5 ± 2.35) and migration (46.14 ± 2.26; P < 0.01 vs. control 21.82 ± 1.62) of DU‐145 +/− cells. In contrast, DU‐145 +/+ cells had lost its response to HGF/SF induced invasion (22.33 ± 2.08; P > 0.05 vs. control 23.5 ± 2.11) and migration (24.12 ± 0.86; P > 0.05 vs. control 23.27 ± 0.81). Conclusions Targeting the HGF/SF receptor by way of a hammerhead ribozyme encoding antisense to c‐Met, is an effective method to reduce the invasive or migration potential in prostate cancer, and may have important therapeutic implications. © 2004 Wiley‐Liss, Inc.