Inhibition of invasion, gelatinase activity, tumor take and metastasis of malignant cells by N‐acetylcysteine

Abstract

The thiol N-acetylcysteine (NAC) is currently considered one of the most promising cancer chemopreventive agents by virtue of its multiple and coordinated mechanisms affecting the process of chemical carcinogenesis. Recent studies have shown that an unpaired cysteine residue in the propeptide plays a key role in inactivation of latent metastasis-associated metalloproteinases: the present study was designed to assess whether NAC could also affect tumor take, invasion and metastasis of malignant cells. As assessed by zymographic analysis, NAC completely inhibited the gelatinolytic activity of type-IV collagenases in the cells tested (gelatinases A and B). Moreover, NAC was efficient in inhibiting the chemotactic and invasive activities of tumor cells of human (A2058 melanoma) and murine origin (K1735 and B16-F10 melanoma cells as well as C87 Lewis lung carcinoma cells) in Boyden-chamber assays, which are predictive of the invasive and metastatic properties. Reduced glutathione (GSH) had a similar, although less effective activity. The number of lung metastases decreased sharply when B16-F10 murine melanoma cells, injected i.v. into nude mice, were pre-treated with NAC and resuspended in medium supplemented with 10 mM NAC. In other experiments NAC was given in drinking water, starting 48-72 hr before subcutaneous inoculation of either B16-F10 cells or of their highly metastatic variant B16-BL6, or intramuscular injection of LLC cells. In all experiments NAC treatment decreased the weight of the locally formed primary tumor and produced a dose-related delay in tumor formation. Spontaneous metastasis formation by B16-F10 and B16-BL6 tumors was slightly yet significantly reduced by oral administration of NAC. However, this was not observed for Lewis lung tumors. These data indicate that NAC affects the process of tumor-cell invasion and metastasis, probably due to inhibition of gelatinases by its sulfhydryl group, with the possible contribution of other mechanisms, including the potent antioxidant activity of this thiol.

Accumulation of hydrogen peroxide is an early and crucial step for paclitaxel‐induced cancer cell death both in vitro and in vivo

“…48 It was proposed that NAC could prevent invasion of tumor cells by inhibiting type IV collagenase. 49 The discrepancy between these results and ours could be explained by differences in tumor cell types, route of tumor cell administration and time of NAC administration.…”

Antileukemia effects of xanthohumol in Bcr/Abl-transformed cells involve nuclear factor-κB and p53 modulation

“…After the incubation period, the invasive cells that migrated into the lower chamber were fixed, stained, and counted under a light microscope. Supernatants of cells from the invasion assay were centrifuged to remove particulates, the protein content was measured by the Bradford method (Bio-Rad), and gelatin zymography was done on identical sample amounts as described previously (12). Gels were then stained in 0.1% Coomassie brilliant blue followed by destaining.…”

In vitro irradiation of basement membrane enhances the invasiveness of breast cancer cells

“…Samples were applied on a 12% polyacrylamide-sodium dodecyl sulfate (SDS) gel containing 0.1% gelatine and electrophoresed at 150 V during 3 h at 41C. After removal of SDS from the gel by incubating in 2.5% Triton X-100 (30 min, four times), the gel was incubated at 371C for 18 h in 40 mM Tris-HCl, pH 7.5, containing 10 mM CaCl 2 , 1 mM ZnCl 2 , 200 mM NaCl and stained with Coomassie blue R-250 (Albini et al, 1995). As positive control, MMP-2 and -9 obtained from Calbiochem were activated with 1 mM APMA as previously described (Paquette et al, 2003) and analysed by zymography.…”