A hammerhead ribozyme suppresses expression of hepatocyte growth factor/scatter factor receptor c-MET and reduces migration and invasiveness of breast cancer cells.

Abstract

PURPOSE:

Hepatocyte growth factor/scatter factor (HGF/SF), via its receptor c-MET, has been implicated to play a pivotal role in breast cancer development and progression. This study examined a transgene-consisting of a combination of U1snRNA, hammerhead ribozyme, and antisense, designed to inhibit c-met expression-and its impact on the migration and in vitro invasion of breast cancer cells.

EXPERIMENTAL DESIGN:

A hammerhead ribozyme targeting human c-MET was cloned into a modified pZeoU1EcoSpe vector and transfected into breast cancer cells MDA MB 231 and MCF-7 by electroporation. Expression of MET mRNA and protein was determined. Migration and in vitro invasiveness of transfected cells were also analyzed.

RESULTS:

Breast cancer cells were transfected with the ribozyme-containing plasmids. Stable transfectants manifested an almost complete loss of MET mRNA and protein, as shown by reverse transcription-PCR, Northern blotting, and Western blotting, respectively, whereas the wild-type plasmid had no effects. Met-ribozyme transfected cells exhibited reduced migration and in vitro invasiveness through extracellular matrix (Matrigel), compared with the wild-type cells and cells transfected with empty plasmid.

CONCLUSIONS:

These data show that targeting c-MET by way of a hammerhead ribozyme encoding antisense to c-MET is an effective approach in reducing the invasiveness of breast cancer cells.

• Authors: W G Jiang, D Grimshaw, J Lane, T A Martin, R Abounader, J Laterra, R E Mansel, R Abounder
• Journal: Clinical cancer research : an official journal of the American Association for Cancer Research (Clin Cancer Res) Vol. 7 Issue 8 Pg. 2555-62 (Aug 2001) ISSN: 1078-0432 [Print] United States
• PMID: 11489839 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Antisense
• RNA, Catalytic
• RNA, Messenger
• RNA, Small Nuclear
• U1 small nuclear RNA
• Proto-Oncogene Proteins c-met

Topics

• Base Sequence
• Breast Neoplasms (genetics, pathology, therapy)
• Cell Movement (genetics)
• DNA, Antisense (genetics)
• Gene Expression Regulation, Neoplastic
• Humans
• Molecular Sequence Data
• Neoplasm Invasiveness (genetics, prevention & control)
• Plasmids (genetics)
• Proto-Oncogene Proteins c-met (genetics, metabolism)
• RNA, Catalytic (genetics, metabolism)
• RNA, Messenger (genetics, metabolism)
• RNA, Small Nuclear (genetics, metabolism)
• Time Factors
• Transfection
• Tumor Cells, Cultured