Inhibition of non‐small cell lung cancer cell migration by grape seed proanthocyanidins is mediated through the inhibition of nitric oxide, guanylate cyclase, and ERK1/2

Abstract

Tumor cell migration is considered as a major event in the metastatic cascade. Here we examined the effect of grape seed proanthocyanidins (GSPs) on migration capacity and signaling mechanisms using nonsmall cell human lung cancer cells. Using in vitro migration assay, we found that treatment of A549 and H1299 cells with GSPs resulted in concentration-dependent inhibition of migration of these cells. The migration capacity of cells was reduced in presence of N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase. GSPs suppressed the elevated levels of endogenous NO/NOS in A549 and H1299 cells and blocked the migration promoting capacity of L-arginine. Treatment with guanylate cyclase (GC) inhibitor 1-H-[1,2,4]oxadiaxolo[4,3-a]quinolalin-1-one (ODQ) reduced the migration of A549 cells whereas additional presence of 8-bromoguanosine 3'5'-cyclic monophosphate (8-Br-cGMP, cGMP analogue) restored the migration of these cells, suggesting a role for GC in migration of A549 cells. GSPs reduced the elevated levels of cGMP in cancer cells and also blocked the migration restoring activity of 8-Br-cGMP. The mitogen-activated protein kinase kinase (MAPKK) inhibitor, UO126, inhibited the migration of A549 cells, indicating a role for MAPKK in the migration. Additionally, UO126 and ODQ inhibited the migration restoring effects of L-arginine in L-NAME-treated cells, suggesting the involvement of cGMP and MAPK pathways in NO-mediated migration. GSPs inhibited L-arginine and 8-Br-cGMP-induced activation of ERK1/2 in A549 cells. Together, these results indicate sequential inhibition of NO/NOS, GC, and MAPK pathways by GSPs in mediating the inhibitory signals for cell migration, an essential step in invasion and metastasis.

Grape Seed Proanthocyanidins Inhibit the Growth of Human Non-Small Cell Lung Cancer Xenografts by Targeting Insulin-Like Growth Factor Binding Protein-3, Tumor Cell Proliferation, and Angiogenic Factors

“…We reported recently that in vitro treatment of NSCLC cells with GSPs inhibited the migration of lung cancer cells (12), which was associated with the inhibition of the levels of nitric oxide production and inducible nitric oxide synthase, guanylate cyclase, and extracellular signal-regulated kinase 1/2. The migration of cancer cells is considered to be a major event in the metastatic cascade of cancer cells.…”

The DDAH/NOS pathway in human prostatic cancer cell lines: Antiangiogenic effect of L-NAME

“…NOS inhibitors have been suggested as antitumor therapeutics (52,65,69,(71)(72)(73)(74)(75)(76)(77)(78)(79)(80)(81). However, the antitumor and antimetastatic effects of NOS inhibitors may be attributed in part to reduced tumor cell invasiveness (52,72,73,78,81) and in part to reduced neovascularization (69,71,74,75,77,79). The importance of the DDAH/NOS pathway in the angiogenic process is confirmed by treatment with the NOS inhibitor L-NAME, which, under our experimental conditions, resulted in a reduction, not only in iNOS and eNOS activity, but also in VEGF, iNOS and eNOS expression.…”

Honokiol, a phytochemical from Magnolia spp., inhibits breast cancer cell migration by targeting nitric oxide and cyclooxygenase-2

“…Cell migration assay. The in vitro migration capacity of breast cancer cells and the inhibitory or stimulatory effects of any agent were determined using Boyden chambers in which two chambers were separated with Millipore membranes (8-μm pore size), as described previously (31). Briefly, cancer cells (1x10 4 cells/100 μl serum-reduced medium) were placed in the upper chamber of a Boyden apparatus.…”