NK-lysin Inhibits Human Hepatoma Cell Invasion And Metastasis Through Modulation Of The Wnt/β-catenin Pathway

Objective we analysed the molecular mechanism of porcine NK-Lysin of our research group in dependent design and expression in vitro on SMMC-7721 invasion and metastasis through modulation of the Wnt/p-catenin pathway.Method Through MTT detection, the safety concentration of NK-Lysin to SMMC-7721 was picked out. Then we divided into five groups, the control group was only added culture medium, the PBS group was added PBS buffer solution and culture medium, the NK-Lysin treated group was added 40μg/mL,20μg/mL and 10μg/mL NK-Lysin buffer. By using wound-healing assay, adhesion assay, invasion assay, the invasion and metastasis of NK-Lysin on SMMC-7721 was detected. By using real time PCR and western blot, the mRNA and protein expressions of β-catenin and MMP-2 were detected.Results ① The results of wound-healing assay showed that, compared with the control group, the migrations in different NK-Lysin group with various concentration were significantly different (p<0.001), while there was no significant difference in PBS group of the migrations (p>0.05). ② The results of adhesion assay showed that, compared with the control group, the adhesion in different NK-Lysin group with various concentration were significantly different (p<0.001), while there was no significant difference in PBS group of the adhesion (P>0.05). ③The results of invasion assay showed that, compared with the control group, the invasion in different NK-Lysin group with various concentration were significantly different 40μg/mL and 20μg/mL (p<0.001), 10μg/mL (p<0.01). while there was no significant difference in PBS group of the invasion (p>0.05). ④The results of real time PCR showed that, compared with the control group, the mRNA expressions of p-catenin and MMP-2 in different NK-Lysin group with various concentration were significantly different. Respectively,40μg/mL (p<0.001),20μg/mL (p<0.01), 10μg/mL (p<0.05)and (p>0.05). While there was no significant difference in PBS group of the mRNA expressions of P-catenin and MMP-2 (p>0.05). ⑤ The results of western blot showed that, compared with the control group, the protein expressions of β-catenin and MMP-2 in different NK-Lysin group with various concentration were significantly different. Respectively,40μg/mL (p<0.001),20μg/mL (p<0.01) and (p<0.05), 10μg/mL (p<0.05) and (p<0.01). while there was no significant difference in PBS group of the protein expressions of β-catenin and MMP-2 (p>0.05).Conclusion Cationic Antimicrobial Peptide NK-Lysin have a inhibition in vitro invasion and metastasis of SMMC-7721 cells. On the other hand, Cationic Antimicrobial Peptide NK-Lysin may be inhibits invasion and metastasis of hepatocellular carcinoma SMMC-7721 through Wnt/p-catenin Signaling Pathway.