[Construction, expression and cell growth inhibition of translocating peptide/granzyme B fusion protein gene].

Aim: To investigate the inhibitory effect of translocating peptide/granzyme B fusion protein on cell growth.

Methods: The translocating peptide gene of Pseudomonas exotoxin A (PE) was fused with active granzyme B gene by recombinant PCR to construct PE II-GrBa fusion protein gene. PE II-mGrBa with a mutation of serine to cystein at active center of GrB was used as negative control. The resulting PE II-GrBa and PE II-mGrBa genes were transiently transfected into mammalian cells via lipofectamine mediation. The effects of expression of PE II-GrBa gene on morphology and growth of transfected cells were detected by MTT colorimetry, TUNEL assay and indirect immunofluorescence staining.

Results: Transient expression of PE II-GrBa resulted in cytoskeleton abnormality, cell growth inhibition, and apoptosis in some cells.

Conclusion: Expression of PE II-GrBa fusion protein can inhibit cell growth.