Elevation of breast carcinoma Nm23-H1 metastasis suppressor gene expression and reduced motility by DNA methylation inhibition

We hypothesize that elevation of Nm23-H1 expression in micrometastatic brea st cancer cells may inhibit their metastatic colonization and further invas ion, and induce differentiation, thus resulting in a clinical benefit. The current study investigated the possible contribution of DNA methylation to the regulation of Nm23-H1 expression, based on the observation that two CpG . islands are present in its promoter. 5-Aza-2'-deoxycytidine (5-Aza-CdR), a DNA methylation inhibitor, increased the Nm23-H1 expression of 5 of 11 hu man breast carcinoma cell lines in vitro, including 3 of 3 metastatically c ompetent lines. Increased Nm23-H1 expression was accompanied by a reduction in motility in vitro, with minimal effect on proliferation. Both increased Nm23-H1 expression and decreased motility were observed using low (75 nM) concentrations of 5-Aza-CdR, Array analysis of MDA-MB-231 breast carcinoma cells treated with 5-Aza-CdR confirmed the elevation of nm23-H1 mRNA, where as relatively few other genes exhibited altered expression, Bisulfite seque ncing of the two CpG islands in a panel of cell lines and in 20 infiltratin g ductal carcinomas revealed that one island (-3090 bp to -3922 bp) exhibit ed infrequent differential methylation. The data indicate that DNA methylat ion inhibitors ran directly or indirectly cause both elevation of Nm23-H1 e xpression and decreased function in one aspect of metastasis, motility.