Mel-18, a polycomb group protein, negatively regulates INK4a/ARF-independent cell cycle progression via inactivation of the Akt signaling pathway in human breast cancer

Abstract

Mel-18, a polycomb group (PcG) protein, has been suggested as a tumor suppressor in human breast cancer. Previously we reported that Mel-18 has anti-proliferative activity in breast cancer cells. However, its functional mechanism has not been fully elucidated. Here, we investigated the role of Mel-18 n human breast cancer. We saw an inverse correlation between Mel-18 and phospho-Akt, which were expressed at low and high levels, respectively, in primary breast tumor tissues from 40 breast cancer patients. The effect of Mel-18 on cell growth was examined in two breast cancer cell lines SK-BR-3 and T-47D, which express relatively low and high levels of endogenous Mel-18, respectively. Upon Mel-18 overexpression in SK-BR-3 cells, cell growth was attenuated and G1 arrest was observed. Likewise, suppression of Mel-18 by antisense expression in T-47D cells lead to enhanced cell growth and accelerated G1/S phase transition. In these cells, Cdk4 and Cdk2 activities were affected by Mel-18, which were mediated by changes in Cyclin D1 expression and p27Kip1 phosphorylation at Thr157, but not by INK4a/ARF genes. The changes were mediated cytoplasmic localization of p27Kip1 was enhanced by Mel-18 suppression in T-47D cells. Akt-mediated cytoplasmic localization of p27Kip1 was inhibited by Mel-18 in SK-BR-3 cells. Moreover, Mel-18 overexpression showed reduced GSK-3 phosphorylation, -catenin nuclear localization, TCF/LEF promoter activity, and cyclin D1 mRNA level. Taken together, we established a linear relationship between Mel-18 ` Akt ` G1 phase regulators.