Ajab Khan¹, Hongquan Li², Sun Na², Panpan Sun², Babar Maqbool³, Asghar Khan¹, Qasim Ali¹, Rahman Ullah¹, Kouhai Fan⁴*

¹Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Swat, Pakistan

²Shanxi Key Lab for Modernization of TCVM, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, PR China

³Faculty of Veterinary and Animal Sciences, University of Agriculture, Dera Ismail Khan, Pakistan

⁴Laboratory Animal Center, Shanxi Agricultural University, Taigu, Shanxi 030801, PR China

 

*Corresponding author’s email: fkhyxj@163.com

Received: 13 January 2026 / Revised: 16 April 2026 / Accepted: 27 April 2026 / Published Online: 18 May 2026

 

Abstract

 

In our previous study, maximum non-toxic concentration (MNTC) of recombinant porcine natural killer lysin (rpNK-Lysin) significantly down-regulated Fascin-1, a prognostic and metastatic biomarker, yet the relevant molecular mechanisms that inhibit Fascin1, was not fully understood. In this study, three different types of hepatocellular carcinoma cell lines (SMMC-7721, MHCC 97H and HepG2) were treated with rpNK-Lysin. Scanning electron microscopy was performed to check its effect on filopodia formation, and the expression of ERK, RSK2, CREB1 and Fascin-1 were determined using qPCR and western blot. Our results showed that MNTC rpNK-lysin successfully down-regulated pERK1/2 which further suppressed the phosphorylation of RSK2 and the transcription factor CREB1, leading to inhibit the CREB transcriptional target Fascin-1, which is involved in filopodia formation. This study confirmed that MNTC rpNK-lysin inhibited metastatic biomarker Fascin1 by down regulating ERK1/2 dependent RSK2 and transcription factor, which further suppressed Fascin1.

 

Keywords: Fascin1, rpNK-lysin, Metastatic biomarker, Hepatocellular carcinoma, Filopodia

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