Background: Some bioactive peptides derived from natural resources or synthesized by rational design
have been shown to have very good anticancer effects. We designed an anticancer fusion peptide (ACFP) based on the
structure of bovine lactoferricin (LfcinB) and hexapeptide (PGPIPN) derived from bovine milk protein.
Objective: To prepare ACFP through genetic engineering and study its antiovarian cancer activity.
Method: ACFP gene was produced by a flexible link arm connecting LfcinB and PGPIPN. ACFP was inductively
expressed in Escherichia coli by the recombinant plasmid pGEX-KG-ACFP. ACFP was prepared and purified by
affinity chromatography, and identified by polyacrylamide gel electrophoresis (PAGE), high-performance liquid
chromatography (HPLC) and mass spectrometry (MS). Recombinant lentivirus vectors were produced by cotransfecting
293T cells with constructed plasmid pLJM1-ACFP, envelope plasmid Δ8.91 and pVSVG using
Lipofectamine. ACFP gene was transfected into ovarian cancer cells by pLJM1-ACFP lentivirus. Cell Viability was
assayed by the methyl thiazolyl tetrazolium (MTT). The apoptosis of ovarian cancer SKOV3 cells was measured by
flow cytometry and observed by Hoechst33258 staining.
Results: ACFP was successfully prepared and purified by genetic engineering. ACFP more effectively inhibited the
viability of human ovarian cancer SKOV3 cells than the single parent peptides in vitro. ACFP was found to have no
cytotoxicity towards untransformed cells. The ACFP gene in cancer cells infected with pLJM1-ACFP lentivirus could
significantly inhibit the viability of SKOV3 cells and induce their apoptosis.
Conclusion: ACFP is a potential therapeutic agent for the treatment of ovarian cancer.