Enhanced Production and Therapeutic Evaluation of TP4 Peptide in Pichia pastoris Against Hepatocellular Carcinoma HUH-7 Cells

Year 2024, Volume: 17 Issue: 2, 505 - 522, 31.08.2024

Seda Kilinç , Mert Karaoglan , Mehmet Kuzucu

https://doi.org/10.18185/erzifbed.1500259

Abstract

Tilapia piscidin 4 (TP4), a cationic antimicrobial peptide, is recognized for its diverse biological roles, including antibacterial, wound-healing, and anticancer properties. Herein, the codon-optimized sequence of TP4 peptide was expressed using the pPICZαA expression vector containing the AOX1 promoter, a strong and inducible promoter, in the Pichia pastoris KM71H expression system. Recombinant TP4 peptide was purified by Ni-NTA affinity chromatography. After purification, the anticancer activity of TP4 was assessed in HUH-7 hepatocellular carcinoma cells, and the underlying mechanisms were determined. In the present study, it was demonstrated for the first time that recombinant TP4 displayed strong anticancer activity in the human HUH-7 cell line. The TP4 antimicrobial peptide can be used as a competitive candidate for the treatment of cancer cells due to its anticancer effects.

Keywords

Tilapia piscidin 4, recombinant peptide, Pichia pastoris, HUH-7

Ethical Statement

There are no ethical issues regarding the publication of this study.

Supporting Institution

This project was supported by Health Institutes of Türkiye (TUSEB) with project no. 31705.

Project Number

31705-TUSEB

Pichia pastoris'te TP4 Peptidinin Artırılmış Üretimi ve Hepatoselüler Karsinom HUH-7 Hücrelerine Karşı Terapötik Değerlendirmesi

Abstract

Bir katyonik antimikrobiyal peptiti olan Tilapia piscidin 4 (TP4), anti-bakteriyel, yara iyileştirici ve antikanser özellikleri de dahil olmak üzere çeşitli biyolojik rolüyle tanınmaktadır. Burada, TP4 peptitinin kodon optimizasyonlu dizisi, Pichia pastoris KM71H ekspresyon sisteminde güçlü ve indüklenebilir bir promotor olan AOX1 promotoru altında pPICZA ekspresyon vektörü kullanılarak eksprese edilmiştir. Rekombinant TP4 peptiti, Ni-NTA afinite kromatografisi ile saflaştırılmıştır. TP4'ün saflaştırılmasından sonra TP4'ün antikanser aktivitesi hepatosellüler karsinoma hücrelerinde değerlendirilmiş ve altta yatan mekanizmalar belirlenmiştir. Bu çalışmada ilk kez rekombinant TP4'ün insan HUH-7 hücre hattında güçlü antikanser aktivite gösterdiği belirlenmiştir. TP4 antimikrobiyal peptiti, antikanser etkileri nedeniyle kanser hücrelerinin tedavisinde rekabetçi bir aday olarak kullanılabilir.

Keywords

Tilapia piscidin 4, rekombinant peptit, Pichia pastoris, HUH-7

Project Number

31705-TUSEB

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