Modulation of MMP-2 and −9 secretion by cytokines, inducers and inhibitors in human melanoma A-2058 cells

Abstract

Abstract. Melanoma, an extremely aggressive cancer, causes the most skin cancer-related deaths, due to metastasis to other areas of the body, such as lymph nodes, lungs, liver, brain or bone. It is characterized by high levels of matrix metalloproteinase (MMP)-2 and -9 secretions that degrade the extracellular matrix and basement membrane, allowing cancer cells to spread to distal organs. Various cytokines, mitogens, growth factors, inducers and inhibitors control MMP activities. We investigated the roles of these in regulation of MMP-2 and -9 in human melanoma A-2058 cells. Human A-2058 cells were grown in DMEM supplemented with 15% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were washed with PBS and incubated in serum-free media with phorbol 12-myristate 13-acetate (PMA) at 10, 25, 50 and 100 ng/ml; TNF-α and IL-1β at 0.1, 1, 10 and 25 ng/ml; LPS at 10, 25, 50 and 100 µg/ml; epigallocatechin gallate (EGCG) and doxycycline (Dox) at 10, 25, 50 and 100 µM without and with PMA; a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract without and with PMA at 10, 50, 100, 500 and 1,000 µg/ml; actinomycin D and cyclohexamide at 2 and 4 µM; retinoic acid and dexamethasone at 50 µM. After 24 h the media were removed and analyzed for MMP-2 and MMP-9 by zymography and densitometry. Melanoma A-2058 demonstrated strong expression of MMP-2 and slight expression of MMP-9. PMA at 100 ng/ml showed no effect on MMP-2 secretion but potently upregulated MMP-9 secretion to 400% that of control. TNF-α showed no significant overall effect on expression of MMP-2 but potent dose-dependent increased MMP-9 secretion with 200% of control at 25 ng/ml. IL-1β showed no significant effect on MMP-2 or MMP-9 secretion by A-2058 cells, except at 25 ng/ml where MMP-2 level was reduced by ~40% and MMP-9 secretion ~50%. LPS treatment showed no significant effect on MMP-2 secretion and enhanced MMP-9 secretion up to 25 µg/ml followed by decreased level. EGCG, NM and doxycycline, without and with PMA, downregulated the expression of MMP-2 and MMP-9 in a dose-dependent manner. Actinomycin D, cyclohexamide and retinoic acid had inhibitory effects on MMP-2, while dexamethasone showed slight stimulatory effect on MMP-2 secretion. Our results showed that select cytokines, mitogens and inhibitors modulated A-2058 MMP-2 and MMP-9 expression. They suggest the clinical potential of MMP inhibitors, especially the non-toxic ones, such as the nutrient mixture and its component EGCG in management of melanoma.

Effect of Sulfated Polysaccharide from Undaria pinnatifida (SPUP) on Proliferation, Migration, and Apoptosis of Human Prostatic Cancer

“…Li et al found that reducing the expression of MMP-9 in prostate cancer cells was accompanied by a decrease in cell invasion and migration [16]. Since we found that SPUP-treated cells inhibit the migration of prostate cancer, we hypothesized that SPUP exerts the effect through cancer cell invasion and metastasis-promoting factor MMP-9 [17] and then found that the higher the concentration of SPUP, the lower the expression of MMP-9, indicating that SPUP may suppress cell migration by inhibiting the expression of MMP-9 protein. In terms of induction of apoptosis, DU145 cells were treated with different SPUP concentrations (25, 50, 100, and 200 μg/ml) for 72 h, and the results indicated that SPUP can induce apoptosis in a dose-dependent manner in DU145 cells.…”

Inflammatory Cytokine Pattern Is Sex-Dependent in Mouse Cutaneous Melanoma Experimental Model

“…Recent data have shown that upon melanoma cell lines, TNF and IL-1 showed no significant overall effect on expression of molecules involved in metastatic processes. Nevertheless, arrays of cytokines/chemokines and various regulatory molecules (e.g., matrix metalloproteinases) can modulate this process [ 34 ]. Our previous human reported TNF values [ 15 ] were as well low and, as prior signaled by us, the low registered circulatory levels can account for the multiplexing assays that use specific antibodies for different circulatory molecules.…”

Surfactin from Bacillus subtilis attenuates ambient air particulate matter-promoted human oral cancer cells metastatic potential

“…Degradation of the ECM is an important step in tumor cell invasion. Although various proteases are involved in ECM degradation, MMPs, a family of zinc and calcium-dependent proteolytic enzymes, digest many components of ECM [32]. MMPs are important to cell migration/invasion and metastasis [32].…”

“…Although various proteases are involved in ECM degradation, MMPs, a family of zinc and calcium-dependent proteolytic enzymes, digest many components of ECM [32]. MMPs are important to cell migration/invasion and metastasis [32]. At least more than 20 different MMPs work on multiple substrates including collagen types I, II, III, IV and stromyelin [32,33].…”