The online version of this article (doi:10.1186/s12885-017-3187-7) contains supplementary material, which is available to authorized users.
Y-box binding protein-1 is an evolutionary conserved transcription and translation regulating protein that is overexpressed in various human malignancies, including breast cancer. Despite reports of YB-1 and its association with distant spread of breast cancer, the intrinsic mechanism underlying this observation remains elusive. This study investigates the role of YB-1 in mediating metastasis in highly invasive breast cancer cell lines.
Silencing the YBX1 gene (which encodes the YB-1 protein) by small interfering RNA (siRNA) was performed in MDA-MB-231 and Hs578T breast cancer cell lines, followed by phenotypic assays including cell migration and invasion assays. Gene expression profiling using Affymetrix GeneChip® Human Transcriptome 2.0 array was subsequently carried out in YB-1 silenced MDA-MB-231 cells. Overexpression and silencing of YBX1 were performed to assess the expression of CORO1C, one of the differentially regulated genes from the transcriptomic analysis. A Gaussia luciferase reporter assay was used to determine if CORO1C is a putative YB-1 downstream target. siRNA-mediated silencing of CORO1C and down-regulation of YBX1 in CORO1C overexpressing MDA-MB-231 cells were performed to evaluate cell migration and invasion.
Downregulation of the YB-1 protein inhibited cell migration and invasion in MDA-MB-231 breast cancer cells. Global gene expression profiling in the YBX1 silenced MDA-MB-231 cells identified differential expression of several genes, including CORO1C (which encodes for an actin binding protein, coronin-1C) as a potential downstream target of YB-1. While knockdown of YBX1 gene decreased CORO1C gene expression, the opposite effects were seen in YB-1 overexpressing cells. Subsequent verification using the reporter assay revealed that CORO1C is an indirect downstream target of YB-1. Silencing of CORO1C by siRNA in MDA-MB-231 cells was also observed to reduce cell migration and invasion. Silencing of YBX1 caused a similar reduction in CORO1C expression, concomitant with a significant decrease in migration in Hs578T cells. In coronin-1C overexpressing MDA-MB-231 cells, increased migration and invasion were abrogated by YB-1 knockdown.
It would appear that YB-1 could regulate cell invasion and migration via downregulation of its indirect target coronin-1C. The association between YB-1 and coronin-1C offers a novel approach by which metastasis of breast cancer cells could be targeted and abrogated.
Additional file 1: Table S1. Sequences of primers used for qPCR. (XLSX 12 kb)12885_2017_3187_MOESM1_ESM.xlsx
Additional file 2: Table S2. Non-coding genes that were differentially expressed following YB-1 silencing in MDA-MB-231 cells. (XLSX 10 kb)12885_2017_3187_MOESM2_ESM.xlsx
Additional file 3: Figure S1. a Selected genes were used to validate the microarray data analysis using qPCR and the expression of all of the genes showed consistent patterns when compared to the microarray data. b Protein expression of some differentially expressed genes were screened by Western blotting in YBX1 silenced MDA-MB-231 cells and the representative blot is shown. (TIFF 403 kb)12885_2017_3187_MOESM3_ESM.tif
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- YBX1 gene silencing inhibits migratory and invasive potential via CORO1C in breast cancer in vitro
Jia Pei Lim
Olivia Jane Scully
Boon Huat Bay
- BioMed Central
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