Saji S, Nakashima S, Hayashi S, Toi M, Saji S, Nozawa Y
2nd Department of Surgery, Gifu University School of Medicine.
Jpn J Cancer Res. 1999 Feb;90(2):210-8. doi: 10.1111/j.1349-7006.1999.tb00735.x.
The overexpression of the oncogene product MDM2 is often observed in human breast cancer cells, especially in estrogen receptor (ER)-positive ones. To study the role of MDM2 protein in ER-positive breast cancer, we have established cell lines derived from MCF-7 which stably express increased and decreased levels of MDM2 by transfection of a mammalian expression vector containing human mdm2 cDNA in sense and antisense orientations, respectively. Interestingly, MDM2 overexpression in MCF-7 cells afforded a remarkable growth advantage under estradiol (E2)-supplemented condition. Then, we analyzed the expression of p53, which is an important regulator of growth and the cell cycle. Unexpectedly, the p53 accumulation induced by E2 was remarkably higher in MCF-7 cells stably overexpressing MDM2 than in the parent MCF-7 cells. On the other hand, reduction of MDM2 suppressed the E2-induced increase in p53 protein. Moreover, mdm2 antisense oligonucleotides prevented E2-induced accumulation of p53. In the steady state, the cellular levels of p53 were also correlated with those of MDM2. These interactions are not consistent with the well-known role of MDM2, which acts as a negative regulator for p53 by inhibiting its function and promoting its rapid degradation. These results suggest that MDM2 may regulate the expression of p53 in the steady state and in response to E2 in breast cancer cells, and imply a novel and important role of MDM2 during breast carcinogenesis.
癌基因产物MDM2的过表达在人类乳腺癌细胞中经常可见,尤其是在雌激素受体(ER)阳性的细胞中。为了研究MDM2蛋白在ER阳性乳腺癌中的作用,我们通过分别转染含有正义和反义方向的人mdm2 cDNA的哺乳动物表达载体,建立了源自MCF-7的细胞系,这些细胞系稳定表达水平升高和降低的MDM2。有趣的是,在补充雌二醇(E2)的条件下,MCF-7细胞中MDM2的过表达赋予了显著的生长优势。然后,我们分析了p53的表达,p53是生长和细胞周期的重要调节因子。出乎意料的是,在稳定过表达MDM2的MCF-7细胞中,E2诱导的p53积累明显高于亲本MCF-7细胞。另一方面,MDM2的减少抑制了E2诱导的p53蛋白增加。此外,mdm2反义寡核苷酸阻止了E2诱导的p53积累。在稳态下,p53的细胞水平也与MDM2的水平相关。这些相互作用与MDM2的众所周知的作用不一致,MDM2通过抑制p53的功能并促进其快速降解而作为p53的负调节因子。这些结果表明,MDM2可能在稳态下以及在乳腺癌细胞中对E2的反应中调节p53的表达,并暗示MDM2在乳腺癌发生过程中具有新的重要作用。