Differential proteomic analysis of a highly metastatic variant of human breast cancer cells using two-dimensional differential gel electrophoresis

Abstract

Distant metastasis represents the major lethal cause of breast cancer. To understand the molecular mechanisms of breast cancer metastasis and identify markers with metastatic potential, we established a highly metastatic variant of parental MDA-MB-231 cells (MDA-MB-231HM). Using two-dimensional electrophoresis (2-DE), we performed a proteomic comparison of the two kinds of cells. As much as 51 protein spots were differentially expressed between the selected variant and its parental counterpart in at least 3 experiments. Ten unique proteins were identified using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS), liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), and database searching software. Among them, nine proteins were up-regulated in MDA-MB-231HM cells, including Macrophage-capping protein (CapG), Galectin-1, Chloride intracellular channel protein 1, Endoplasmic reticulum protein ERp29 precursor, Stathmin-1 (STMN1), Isoform 1 of uridine-cytidine kinase 2(UCK2), Rho GDP-dissociation inhibitor 2 (ARHGDIB), isocitrate dehydrogenase [NADP] cytoplasmic (IDH1), and N-myc downstream regulated gene 1 (NDRG1) protein. Only transgelin-2 was down-regulated. Differential expression was confirmed for three proteins including CapG, STMN1, and transgelin-2 by Western blotting analysis. Transgelin-2 was chosen for further verification by immunohistochemistry. The results suggested that 2-DE would be an efficient way to screen the proteins responsible for specific biological function. Furthermore, the findings imply that different proteins may be involved in the metastatic process in breast carcinomas.

A novel interplay between oncogenic PFTK1 protein kinase and tumor suppressor TAGLN2 in the control of liver cancer cell motility

“…It is frequently downregulated in breast, colon (Shields et al, 2002;Wulfkuhle et al, 2002) and prostate cancers (Yang et al, 2007), where a tumor suppressor function has been proposed. In colorectal cancer, negative TAGLN expression has further been reported to be an independent prognostic indicator for shorter patient's survival (Zhao et al, 2009), whereas a role in impeding metastasis was suggested in breast carcinoma (Xu et al, 2010). TAGLN also acts to suppress the expression of metallo-matrix proteinase MMP9, which is an important enzyme involved in the tissue remodeling during cancer cells invasion into the surrounding tissues (Nair et al, 2006).…”

Prognostic Evaluation of CapG, Gelsolin, P‐gp, GSTP1, and Topo‐II Proteins in Non‐Small Cell Lung Cancer

“…CapG overexpression has been reported in several cancer types, such as breast carcinoma (Kang et al, 2010;Xu et al, 2010), ovarian adenocarcinoma (Partheen et al, 2008), oral carcinoma (Nomura et al, 2008), pancreatic cancer (Thompson et al, 2007a,b), and ocular melanoma (Van Ginkel et al, 1998).…”

“…A clinical study of pancreatic ductal adenocarcinomas previously showed that high CapG expression was correlated with increased tumor size (Thompson et al, 2007a,b). Some researchers have reported that RNAi knockdown of CapG in human cancer cells, such as breast and pancreatic cancer cells can significantly reduce the invasion and motility of cancer cells and cell aggression (Dahl et al, 2005;Thompson et al, 2007a,b;Kang et al, 2010;Xu et al, 2010). CapG is considered to be a tumor promoter and can modulate invasive properties of cells during tumorigenesis (Veerle et al, 2004).…”

Inhibition of N-Myc down regulated gene 1 inin vitrocultured human glioblastoma cells

“…NDRG1 protein expression has been described to be present in normal brain or brain tumor tissue [11] as well as being an important gene that is playing an active role in the regulation of a broad spectrum of human cancer diseases like human gastric cancer, squamous cell carcinomas, breast cancer, human hepatocellular carcinoma, brain tumors and leukemia [12][13][14][15][16][17][18][19][20][21][22] . NDRG1 was suggested to be a prognostic marker for hypoxic regions within a tumor mass because of its stability as a protein [23][24][25] and because it is highly expressed in malignant tumor tissues compared to normal tissue of the same origin [26] .…”

“…g/mL streptomycin, and 100 units/mL penicillin). The pSUPER-NDRG1 vectors were designed as mentioned before [10][11][12][13][14][15][16][17] . To establish pSUPER-NDRG1, two sequences were selected from the human NDRG1 cDNA (5'-GCATTATTGGCAT-GGGAAC-3' (positions 398-416) and 5'-ATGCAGAG-TAACGTGGAAG-3' (positions 601 to 619), relative to the start codon).…”