Role of Tumor-Associated Macrophages on Cathepsin-B, Cathepsin-D, MMP-2, and MMP-9 in HSC-3 Oral Cancer Cells

摘要

This study was aimed to investigate the role of macrophages on an expression of cathepsin B and D (CTSB and CTSD) and the production of matrix metalloproteinase 2 and 9 (MMP-2 and 9) in lymph node metastatic human oral cancer, HSC-3 cell line. Human monocytic cell line (THP-1) was differentiated into 3 types of macrophages; M0, M1 and M2 macrophage using phorbol 12-myristate 13-acetate (PMA), Porphyromonas gingivalis lipopolysaccharide (Pg LPS) and interleukin-4 (IL-4), respectively. Total RNA was extracted from THP-1 and macrophages to detect the expression of CD14, CD68 and CD206 by real-time PCR. The non-contact co-culture of HSC-3 with conditioned media (CM) from different phenotype of macrophages was performed. After 24 hours, the expression of CTSB and CTSD in HSC-3 was investigated using real-time PCR. At 48 hours of coculture, MMP-2 and MMP-9 activities were detected using gelatin zymography. We successfully converted THP-1 into M0, M1 and M2 macrophages. Co-culture of HSC-3 with CM from M2 macrophage significantly elevated the expressions of CTSB and CTSD (p < 0.05). Co-culture of HSC-3 with CM from M2 macrophages also increased MMP-9 activity (p < 0.05) but not MMP-2 activity was found no difference. Hence, M2 macrophages could elevate the gene expression of CTSB and CTSD and the production of MMP-9 in HSC-3, and may play role in invasion, and metastasis of oral cancers.