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MRNA and lncRNA expression profiles in MCF-7 breast cancer cells stably overexpressing pcDNA3.1-PinX1 and Empty Vector

ABSTRACT: To elucidate the molecular mechanism and signaling pathways that PinX1 is involved in, a Human LncRNA Array V2.0 (Arraystar, USA)-based genome wide screen of the alterations in lncRNA and mRNA expression profiles was performed in MCF-7 cells stably overexpressing PinX1. MCF-7 breast cancer cells were transfected with the pcDNA3.1-PinX1 and pcDNA3.1 empty vector and the microarray analysis were performed after the clonal selection of cells with a high expression level.

ORGANISM(S): Homo sapiens

SUBMITTER: Shi Rong

PROVIDER: S-ECPF-GEOD-46756 | biostudies-other |

REPOSITORIES: biostudies-other

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Project description:Mitochondrial ferritin (FtMt) is an iron-transport protein with ferroxidase properties localized to mitochondria. Levels are generally low in all tissues, while increasing the expression of FtMt in neuronal-like cells has been shown to be protective. To determine whether FtMt has potential as a therapeutic approach, there remains the question of how much FtMt is protective. To address this issue, we transfected SH-SY5Y neuroblastoma cells with a FtMt expression plasmid and isolated cell lines with stable expression of FtMt at high, medium and low levels. Using these cell lines, we examined effects of FtMt on neuronal phenotype, neuroprotective activity and gene expression profiles. The phenotypic properties of high, medium and low FtMt expressors were compared with native untransfected SH-SY5Y cells after differentiation with retinoic acid to a neuronal phenotype. Overexpression of FtMt, even in low expressing cells, showed significant protection from oxidative stress induced by hydrogen peroxide or cobalt chloride. Higher levels of FtMt expression did not appear to offer greater protection, and did not have toxic consequences to cells, even though there were significantly more aggregated mitochondria in the highest expressing clone. The phenotypes differed between cell clones when assessed by cell growth, neurite outgrowth, and expression of neuronal proteins including those associated with neurodegenerative diseases. Microarray analysis of high, medium and negative FtMt-expressing cells identified different patterns of expression of certain genes associated with oxidative stress and neuronal development, amongst others. Validation of microarray analyses was carried out by real time polymerase chain reaction. The results showed significant differences in expression of thioredoxin-interacting protein (TXNIP) and microsomal glutathione transfer-1 (MGST-1), which can have critical roles in the regulation of oxidative stress. Differences in expression of calcitonin-related polypeptide alpha (CALCA), growth differentiation factor-15 (GDF-15) and secretogranin II (SCG2) were also observed. Our findings indicate that even low levels of increased FtMt expression can be protective possibly by alterations of some oxidative stress-related and growth factor genes, while high levels of expression did not appear to offer greater protection from oxidative stress or induce significant toxicity in cells. These experiments provide supporting data that increasing FtMt might be a feasible strategy for therapeutics in certain neurodegenerative and neurological diseases.

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MRNA and lncRNA expression profiles in MCF-7 breast cancer cells stably overexpressing pcDNA3.1-PinX1 and Empty Vector

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