Differentially expressed proteins in ER+ MCF7 and ER- MDA- MB-231 human breast cancer cells by RhoGDI-α silencing and overexpression.

BACKGROUND: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDIα) activity on migration and invasion of estrogen receptor positive (ER+) and negative (ER-) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDIα and other proteins interacting directly or indirectly with RhoGDIα in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. MATERIALS AND METHODS: ER+ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/time- of-flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDIα using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDIα. RESULTS: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDIα in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDIα in MCF7, while only one protein was identified in the upregulation of RhoGDIα in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-α activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. CONCLUSIONS: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDIα with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration.

Downregulation of RhoGDIα increased migration and invasion of ER (+) MCF7 and ER (-) MDA-MB-231 breast cancer cells.

Rho GDP dissociation inhibitors (RhoGDIs) can inhibit cell motility, invasion, and metastasis in cancer by inactivating the RhoGTPases. A member of RhoGDI family has been consistently shown to interact with estrogen receptor (ER), and change its transcriptional activity. ER is a receptor known to be inversely correlated with cell motility and invasion in breast cancer. The consequence of RhoGDIα activity on migration and invasion of ER (+) and ER (-) breast cancers is not clear. The aim of our study was to investigate the possible opposing effect of RhoGDIα on the migration and invasion of ER (+) MCF7 and ER (-) MDA-MB-231 breast cancer cells. RhoGDIα was downregulated using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDIα, and their ability for migration and invasion was assayed using transwell chambers. It was found that the silencing of RhoGDIα in MCF7 and MDA-MB-231 cells significantly increased migration and invasion of these cells into the lower surface of porous membrane of the chambers. Overexpression of RhoGDIα in MCF7 cells suppressed their migration and invasion, but no significant effect was found on MDA-MB-231 cells. Our results indicate that the downregulation of RhoGDIα similarly affects the in vitro migration and invasion of ER (+) MCF7 and ER (-) MDA-MB-231 cells. However, our assays are differently affected by the upregulation of RhoGDIα in these two cell lines and this may be due to the differences in ER expression, primary invasive ability and/or other molecules between these two cell line models which warrant further investigation.