Background & Objective: Invasion and metastasis are significant characteristics of cancer cells, Type IV collagenase (matrix metalloproteinase-2, and-9) plays an important role in cancer invasion and metastasis. This study was designed to block the secretion of type IV collagenase via intracellular antibody (intrabody) methods, and inhibit cancer invasion and metastasis.

Methods: We constructed expression plasmid pcDNA3.1-ER.scFv coding for an endoplasmic reticulum (ER)- retained, single chain antibody (ER.scFv) against the type IV collagenase. The intrabody gene was transfected into human giant cell pulmonary carcinoma PG cells. Western blot was performed to detect the expression of pcDNA3.1-ER.scFv. Co-immunoprecipitation was used to analyse the interaction between ER. scFv and target protein in PG cells. The secretion of type IVcollagenase was detected by gelatin zymography. Cell behavior was examined by invasion and proliferation assay in vitro.

Results: ER. scFv expressed in PG cells, and can recognize and combine its cognate target protein matrix metalloproteinase-9. Intrabody gene transfection significantly blocked the function and activity of type IV collegenase. The ER.scFv-transfected PG cells were less invasive than parental or vector control cells, the suppression rate of ER.scFv-transfected PG cells was 76.3%(P< 0.05). ER. scFv showed anti-proliferative effect on PG cells cultured on Matrigel.

Conclusions: ER-retained intrabody technology may restrain the activity of type IV collagenase in protein processing and secretory pathway, and inhibits cancer cell invasion and proliferation. Anti-type IV collagenase intrabody may be further used in cancer gene therapy.

Download full-text PDF

Source

Publication Analysis

Top Keywords

type collagenase
16
collagenase intrabody
12
invasion proliferation
12
anti-type collagenase
8
invasion metastasis
8
cells
8
cancer invasion
8
secretion type
8
intrabody gene
8
target protein
8

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!