Enhancement of Anti-tumor Effect of a Ribosome-inactivating Protein by Cell Penetrating Peptides and Saponin

doi:10.13560/j.cnki.biotech.bull.1985.2019-0198

Abstract

Abstract: A ribosome-inactivated protein named as Burkholderia lethal factor 1（BLF1）was fused with the cell penetrating peptide（CPP）HBP and co-administered with esculentoside A（EsA）to enhance the antitumor effect of the recombinant protein BLF1-HBP. Both the recombinant protein BLF1 and BLF1-HBP were expressed in Escherichia coliBL21（DE3）and then purifed with NI-NTA. The inhibitory effects of the two recombinant proteins were tested and compared in tumor cell line HepG2,MCF-7,A549 and HeLa with the MTT assay. The transmembrane efficiency of the recombinant proteins was observed by laser confocal microscopy and the anti-tumor effect was analyzed with flow cytometry. The result showed that CPP HBP effectively increased the inhibitory effect of BLF1 in all 4 tested tumor cell lines,and the most significant inhibitory effect was observed in HeLa cells,which was increased by 47.5 times. When BLF1-HBP was co-administered with EsA,the antitumor activity of BLF1-HBP was further improved and the IC₅₀ value in MCF-7 cells decreased from 6 840 nmol/L to 0.57 nmol/L. Laser confocal observation revealed that EsA effectively promoted the translocation efficiency of BLF1 recombinant protein. Flow cytometry analysis showed that EsA greatly enhanced the apoptosis effect of tumor cells induced by BLF1-HBP. Therefore,the fusion of BLF1 and HBP and co-administration with EsA may significantly increase the antitumor effect of BLF1,and this enhancement is due to the induction of more tumor cell apoptosis.

Key words: Burkholderia lethal factor 1, cell penetrating peptide, esculentoside A, antitumor effect

LIU Yang, CAO Xue-wei, LU Mei-ya, WANG Fu-jun, ZHAO Jian. Enhancement of Anti-tumor Effect of a Ribosome-inactivating Protein by Cell Penetrating Peptides and Saponin[J]. Biotechnology Bulletin, 2019, 35(8): 146-154.

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