Construction of an Anticancer Fusion Peptide (ACFP) Derived from Milk Proteins and an Assay of Anti-ovarian Cancer Cells in vitro

Author(s): Juan Zhou, Xue Yang, Wenxiao Zhang, Jing Wang, Cai Wei, Fang Gu, Ting Lei, Yide Qin*.

Journal Name: Anti-Cancer Agents in Medicinal Chemistry

Volume 17 , Issue 4 , 2017

Submit Manuscript
Submit Proposal

Abstract:

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.

Keywords: Fusion peptide, construction, preparation, lentivirus, anticancer, genetic engineering, apoptosis, therapeutic agent.

Rights & PermissionsPrintExport Cite as


Article Details

VOLUME: 17
ISSUE: 4
Year: 2017
Page: [635 - 643]
Pages: 9
DOI: 10.2174/1871520616666160627091131
Price: $58

Article Metrics

PDF: 25
HTML: 1