Hooshmand Somayeh, Ghaderi Abbas, Yusoff Khatijah, Thilakavathy Karuppiah, Rosli Rozita, Mojtahedi Zahra
Department of Obstetrics and Gynaecology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Selangor, Malaysia E-mail :
Asian Pac J Cancer Prev. 2014;15(7):3311-7. doi: 10.7314/apjcp.2014.15.7.3311.
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.
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.
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.
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.
尚未使用蛋白质组学方法研究Rho GDP解离抑制剂α（RhoGDIα）活性对雌激素受体阳性（ER +）和阴性（ER -）乳腺癌细胞迁移和侵袭的影响。RhoGDIα以及其他与RhoGDIα直接或间接相互作用的蛋白质在具有不同转移潜能的MCF7和MDA-MB-231细胞中的表达变化尤其令人关注。
ER + MCF7和ER - MDA-MB-231细胞系进行二维电泳（2-DE），使用短干扰RNA（siRNA）下调RhoGDIα并使用RhoGDIα的绿色荧光蛋白标记的开放阅读框（ORF）克隆上调后，通过基质辅助激光解吸/电离飞行时间/飞行时间（MALDI-TOF/TOF）质谱（MS）分析鉴定感兴趣的斑点。
结果显示这些细胞中共有35种蛋白质上调或下调。在这里，我们分别在MCF-7和MDA-MB-231细胞中鉴定出9种和15种随着RhoGDIα沉默而差异表达的蛋白质。此外，在MCF7中RhoGDIα上调时有10种蛋白质差异表达，而在MDA-MB-231中RhoGDIα上调时仅鉴定出一种蛋白质。基于这些蛋白质的生物学功能，结果表明参与细胞迁移的蛋白质受RhoGDI-α活性的影响更大。虽然这些蛋白质中的几种先前已被指出与乳腺癌细胞的肿瘤发生和侵袭有关，但有些以前尚未报道参与乳腺癌迁移。因此，这些蛋白质可能是乳腺癌细胞肿瘤发生和侵袭的有用候选生物标志物。
需要进一步的研究来确定这些蛋白质调节细胞迁移的机制。RhoGDIα与其他潜在生物标志物的联合应用可能是抑制乳腺癌细胞迁移更有前景的方法。
