α-Lipoic acid suppresses migration and invasion via downregulation of cell surface ß1-integrin expression in bladder cancer cells.

Our previous study showed α-lipoic acid (LA) downregulated cell surface ß1-integrin expression of v-H-ras-transformed derivative of rat fibroblast with amelioration of their malignant phenotype. Here, we evaluated the ameliorating effect of LA on the malignant characters in H-ras-transformed bladder cancer cells. H-ras mutated bladder cancer line, T24 cells were incubated with LA to evaluate the inhibitory effect on proliferation, migration, invasion and ß1-integrin expression. Fluorescence staining of F-actin and western blotting analyses of the related signaling pathways were also performed. LA inhibited the proliferation of T24 cells. Cell adhesion to collagen IV and fibronectin was strikingly inhibited by LA treatment accompanied by downregulation of cell surface but not whole cell ß1-integrin expression. LA clearly inhibited cell migration and invasion of T24 cells, which were mimicked by extracellular signal-regulated kinase (ERK) and Akt pathway inhibition. Actually, LA significantly downregulated the phosphorylated ERK and Akt levels. Moreover, LA downregulated phosphorylated focal adhesion kinase level with disappearance of stress fiber formation. Finally, although LA induced the internalization of cell surface ß1-integrin, disruption of the raft did not affect the action of LA. Taken together, LA is a promising agent to improve malignant character of bladder cancer cells through regulation of cellular ß1-integrin localization.

Selective inhibition by apocynin of the proliferation and adhesion to fibronectin of v-H-ras-transformed 3Y1 cells.

We determined the effects of apocynin, a representative inhibitor of NADPH oxidase, on the proliferative and adhesive properties of 3Y1 rat fibroblasts and the 3Y1 v-H-ras-transformed derivative, HR-3Y1-2. Apocynin inhibited the proliferation of HR-3Y1-2 but not 3Y1 cells at 10 µM and 100 µM. Apocynin also decreased the intracellular reactive oxygen species (ROS) level in HR-3Y1-2 but not 3Y1 cells. We also evaluated the effects of apocynin on cell adhesion to fibronectin and found decreased adhesion of HR-3Y1-2 cells to fibronectin-coated plates. Our results indicate that apocynin selectively down-regulated ß1-integrin cell surface expression on the HR-3Y1-2 cells. It also inhibited the migration and invasion of these cells. These data suggest that reducing the production of NADPH oxidase-mediated ROS could be an effective means for ameliorating the abnormal growth, adhesion and motility of v-H-ras-transformed cells.

Alpha lipoic acid selectively inhibits proliferation and adhesion to fibronectin of v-H-ras-transformed 3Y1 cells.

Here, we focused on the effects of racemic α-lipoic acid on proliferation and adhesion properties of 3Y1 rat fibroblasts and the v-H-ras-transformed derivative, HR-3Y1-2 cells. Racemic α-lipoic acid inhibited proliferation of HR-3Y1-2 but not 3Y1 cells at 0.3 and 1.0 mM. R-(+)-α-lipoic acid also inhibited proliferation of HR-3Y1-2 cells equivalent to that of racemic α-lipoic acid. In addition, racemic α-lipoic acid decreased intracellular reactive oxygen species levels in HR-3Y1 cells but not 3Y1 cells. Next, we evaluated the effects of racemic α-lipoic acid on cell adhesion to fibronectin. The results indicated that racemic α-lipoic acid decreased adhesive ability of HR-3Y1-2 cells to fibronectin-coated plates. As blocking antibody experiment revealed that ß1-integrin plays a key role in cell adhesion in this experimental system, the effects of racemic α-lipoic acid on the expression of ß1-integrin were examined. The results indicated that racemic α-lipoic acid selectively downregulated the expression of cell surface ß1-integrin expression in HR-3Y1-2 cells. Intriguingly, exogenous hydrogen peroxide upregulated cell surface ß1-integrin expression in 3Y1 cells. Taken together, these data suggest that reduction of intracellular reactive oxygen species levels by α-lipoic acid could be an effective means of ameliorating abnormal growth and adhesive properties in v-H-ras transformed cells.